Deformation of network connectivity in the inferior olive of connexin 36-deficient mice is compensated by morphological and electrophysiological changes at the single neuron level.
AuthorsDe Zeeuw CI, Chorev E, Devor A, Manor Y, Van Der Giessen RS, De Jeu MT, Hoogenraad CC, Bijman J, Ruigrok TJ, French P, Jaarsma D, Kistler WM, Meier C, Petrasch-Parwez E, Dermietzel R, Sohl G, Gueldenagel M, Willecke K, Yarom Y
JournalJ Neurosci
PubMed ID12805309
Compensatory mechanisms after genetic manipulations have been documented extensively for the nervous system. In many cases, these mechanisms involve genetic regulation at the transcription or expression level of existing isoforms. We report a novel mechanism by which single neurons compensate for changes in network connectivity by retuning their intrinsic electrical ... More
Cyclin G2 associates with protein phosphatase 2A catalytic and regulatory B' subunits in active complexes and induces nuclear aberrations and a G1/S phase cell cycle arrest.
AuthorsBennin David A; Don Aruni S Arachchige; Brake Tiffany; McKenzie Jennifer L; Rosenbaum Heidi; Ortiz Linette; DePaoli-Roach Anna A; Horne Mary C;
JournalJ Biol Chem
PubMed ID11956189
Cyclin G2, together with cyclin G1 and cyclin I, defines a novel cyclin family expressed in terminally differentiated tissues including brain and muscle. Cyclin G2 expression is up-regulated as cells undergo cell cycle arrest or apoptosis in response to inhibitory stimuli independent of p53 (Horne, M., Donaldson, K., Goolsby, G., ... More
Cytoplasmic sequestration of HDAC7 from mitochondrial and nuclear compartments upon initiation of apoptosis.
AuthorsBakin RE, Jung MO
JournalJ Biol Chem
PubMed ID15364908
'Control of global histone acetylation status is largely governed by the opposing enzymatic activities of histone acetyltransferases and deacetylases (HDACs). HDACs were originally identified as modulators of nuclear histone acetylation status and have been linked to chromosomal condensation and subsequent gene repression. Accumulating evidence highlights HDAC modification of non-histone targets. ... More
The scaffold protein gravin (cAMP-dependent protein kinase-anchoring protein 250) binds the beta 2-adrenergic receptor via the receptor cytoplasmic Arg-329 to Leu-413 domain and provides a mobile scaffold during desensitization.
AuthorsFan G, Shumay E, Wang H, Malbon CC
JournalJ Biol Chem
PubMed ID11309381
'The cyclic AMP-dependent kinase-anchoring proteins (AKAPs) function as scaffolds for a wide-range of protein-protein interactions. The 250-kDa AKAP known as gravin plays a central role in organizing G-protein-coupled receptors to the protein kinases and phosphatases that regulate receptor function in desensitization, resensitization, and sequestration. Although gravin is critical for G-protein-linked ... More
Characterization of Mayven, a novel actin-binding protein predominantly expressed in brain.
AuthorsSoltysik-Espanola M, Rogers RA, Jiang S, Kim TA, Gaedigk R, White RA, Avraham H, Avraham S
JournalMol Biol Cell
PubMed ID10397770
'The cytoskeleton plays an important role in neuronal morphogenesis. We have identified and characterized a novel actin-binding protein, termed Mayven, predominantly expressed in brain. Mayven contains a BTB (broad complex, tramtrack, bric-a-brac)/POZ (poxvirus, zinc finger) domain-like structure in the predicted N terminus and "kelch repeats" in the predicted C-terminal domain. ... More
Chromosome and replisome dynamics in E. coli: loss of sister cohesion triggers global chromosome movement and mediates chromosome segregation.
AuthorsBates D, Kleckner N
JournalCell
PubMed ID15960977
'Chromosome and replisome dynamics were examined in synchronized E. coli cells undergoing a eukaryotic-like cell cycle. Sister chromosomes remain tightly colocalized for much of S phase and then separate, in a single coordinate transition. Origin and terminus regions behave differently, as functionally independent domains. During separation, sister loci move far ... More
Syntaxin 7 is localized to late endosome compartments, associates with Vamp 8, and Is required for late endosome-lysosome fusion.
AuthorsMullock BM, Smith CW, Ihrke G, Bright NA, Lindsay M, Parkinson EJ, Brooks DA, Parton RG, James DE, Luzio JP, Piper RC
JournalMol Biol Cell
PubMed ID10982406
'Protein traffic from the cell surface or the trans-Golgi network reaches the lysosome via a series of endosomal compartments. One of the last steps in the endocytic pathway is the fusion of late endosomes with lysosomes. This process has been reconstituted in vitro and has been shown to require NSF, ... More
Tec kinase signaling in T cells is regulated by phosphatidylinositol 3-kinase and the Tec pleckstrin homology domain.
AuthorsYang WC, Ching KA, Tsoukas CD, Berg LJ
JournalJ Immunol
PubMed ID11123316
'Tec, the prototypical member of the Tec family of tyrosine kinases, is abundantly expressed in T cells and other hemopoietic cell types. Although the functions of Itk and Txk have recently been investigated, little is known about the role of Tec in T cells. Using antisense oligonucleotide treatment to deplete ... More
The cohesion protein ORD is required for homologue bias during meiotic recombination.
AuthorsWebber HA, Howard L, Bickel SE
JournalJ Cell Biol
PubMed ID15007062
'During meiosis, sister chromatid cohesion is required for normal levels of homologous recombination, although how cohesion regulates exchange is not understood. Null mutations in orientation disruptor (ord) ablate arm and centromeric cohesion during Drosophila meiosis and severely reduce homologous crossovers in mutant oocytes. We show that ORD protein localizes along ... More
Alexa dyes, a series of new fluorescent dyes that yield exceptionally bright, photostable conjugates.
AuthorsPanchuk-Voloshina N, Haugland RP, Bishop-Stewart J, Bhalgat MK, Millard PJ, Mao F, Leung WY, Haugland RP
JournalJ Histochem Cytochem
PubMed ID10449539
'Alexa 350, Alexa 430, Alexa 488, Alexa 532, Alexa 546, Alexa 568, and Alexa 594 dyes are a new series of fluorescent dyes with emission/excitation spectra similar to those of AMCA, Lucifer Yellow, fluorescein, rhodamine 6G, tetramethylrhodamine or Cy3, lissamine rhodamine B, and Texas Red, respectively (the numbers in the ... More
Distal renal tubular acidosis in mice that lack the forkhead transcription factor Foxi1.
AuthorsBlomqvist SR, Vidarsson H, Fitzgerald S, Johansson BR, Ollerstam A, Brown R, Persson AE, Bergström G G, Enerbäck S
JournalJ Clin Invest
PubMed ID15173882
'While macro- and microscopic kidney development appear to proceed normally in mice that lack Foxi1, electron microscopy reveals an altered ultrastructure of cells lining the distal nephron. Northern blot analyses, cRNA in situ hybridizations, and immunohistochemistry demonstrate a complete loss of expression of several anion transporters, proton pumps, and anion ... More
Evidence that Ca(2+) cycling by the plasma membrane Ca(2+)-ATPase increases the 'excitability' of the extracellular Ca(2+)-sensing receptor.
AuthorsDe Luisi A, Hofer AM
JournalJ Cell Sci
PubMed ID12640037
'The extracellular Ca(2+)-sensing receptor (CaR) is a widely expressed G-protein-coupled receptor that translates information about [Ca(2+)] in the extracellular milieu to the interior of the cell, usually via intracellular Ca(2+) signaling pathways. Using fura-2 imaging of cytoplasmic [Ca(2+)], we observed that HEK293 cells expressing CaR oscillated readily under conditions permissive ... More
Removal of the membrane-anchoring domain of epidermal growth factor leads to intracrine signaling and disruption of mammary epithelial cell organization.
AuthorsWiley HS, Woolf MF, Opresko LK, Burke PM, Will B, Morgan JR, Lauffenburger DA
JournalJ Cell Biol
PubMed ID9832559
'Autocrine EGF-receptor (EGFR) ligands are normally made as membrane-anchored precursors that are proteolytically processed to yield mature, soluble peptides. To explore the function of the membrane-anchoring domain of EGF, we expressed artificial EGF genes either with or without this structure in human mammary epithelial cells (HMEC). These cells require activation ... More
Polar localization of virulence-related Esx-1 secretion in mycobacteria.
AuthorsCarlsson F, Joshi SA, Rangell L, Brown EJ,
JournalPLoS Pathog
PubMed ID19180234
'The Esx-1 (type VII) secretion system is critical for virulence of both Mycobacterium tuberculosis and Mycobacterium marinum, and is highly conserved between the two species. Despite its importance, there has been no direct visualization of Esx-1 secretion until now. In M. marinum, we show that secretion of Mh3864, a novel ... More
Interaction of c-Jun amino-terminal kinase interacting protein-1 with p190 rhoGEF and its localization in differentiated neurons.
AuthorsMeyer D, Liu A, Margolis B
JournalJ Biol Chem
PubMed ID10574993
'c-Jun amino-terminal kinase (JNK) interacting protein-1 (JIP-1) was originally identified as a cytoplasmic inhibitor of JNK. More recently, JIP-1 was proposed to function as a scaffold protein by complexing specific components of the JNK signaling pathway, namely JNK, mitogen-activated protein kinase kinase 7, and mixed lineage kinase 3. We have ... More
Palmitoylation of apolipoprotein B is required for proper intracellular sorting and transport of cholesteroyl esters and triglycerides.
AuthorsZhao Y, McCabe JB, Vance J, Berthiaume LG
JournalMol Biol Cell
PubMed ID10679026
'Apolipoprotein B (apoB) is an essential component of chylomicrons, very low density lipoproteins, and low density lipoproteins. ApoB is a palmitoylated protein. To investigate the role of palmitoylation in lipoprotein function, a palmitoylation site was mapped to Cys-1085 and removed by mutagenesis. Secreted lipoprotein particles formed by nonpalmitoylated apoB were ... More
Phosphatidylinositol 3-kinase, protein kinase C, and MEK1/2 kinase regulation of dopamine transporters (DAT) require N-terminal DAT phosphoacceptor sites.
'The dopamine transporter (DAT) modulates dopamine neurotransmission and is a primary target for psychostimulant influences on locomotion and reward. Selective DAT expression by dopaminergic neurons has led to use of cocaine analog DAT radioligands to assess rates of progression of dopamine neuronal degeneration in Parkinson''s disease. We have documented that ... More
Optimized fixation and immunofluorescence staining methods for Dictyostelium cells.
AuthorsHagedorn M, Neuhaus EM, Soldati T,
JournalMethods Mol Biol
PubMed ID16957300
'Recent years have seen a powerful revival of fluorescence microscopy techniques, both to observe live cells and fixed objects. The limits of sensitivity, simultaneous detection of multiple chromophores, and spatial resolution have all been pushed to the extreme. Therefore, it is essential to improve in parallel the quality of the ... More
Gbeta gamma isoforms selectively rescue plasma membrane localization and palmitoylation of mutant Galphas and Galphaq.
AuthorsEvanko DS, Thiyagarajan MM, Siderovski DP, Wedegaertner PB
JournalJ Biol Chem
PubMed ID11294873
'Mutation of Galpha(q) or Galpha(s) N-terminal contact sites for Gbetagamma resulted in alpha subunits that failed to localize at the plasma membrane or undergo palmitoylation when expressed in HEK293 cells. We now show that overexpression of specific betagamma subunits can recover plasma membrane localization and palmitoylation of the betagamma-binding-deficient mutants ... More
Involvement of nuclear factor of activated T cells activation in UV response. Evidence from cell culture and transgenic mice.
AuthorsHuang C, Mattjus P, Ma WY, Rincon M, Chen NY, Brown RE, Dong Z
JournalJ Biol Chem
PubMed ID10734048
'Mammalian cells respond to UV radiation by signaling cascades leading to activation of transcription factors, such as activated protein 1, NFkappaB, and p53, a process known as the "UV response." Nuclear factor of activated T cells (NFAT) was first identified as an inducible nuclear factor in immune response and subsequently ... More
Differential requirement for classic and novel PKC isoforms in respiratory burst and phagocytosis in RAW 264.7 cells.
AuthorsLarsen EC, DiGennaro JA, Saito N, Mehta S, Loegering DJ, Mazurkiewicz JE, Lennartz MR
JournalJ Immunol
PubMed ID10946313
'The binding of Ab (IgG)-opsonized particles by FcgammaRs on macrophages results in phagocytosis of the particles and generation of a respiratory burst. Both IgG-stimulated phagocytosis and respiratory burst involve activation of protein kinase C (PKC). However, the specific PKC isoforms required for these responses have yet to be identified. We ... More
Relationship between novel isoforms, functionally important domains, and subcellular distribution of CD164/endolyn.
AuthorsChan JY, Lee-Prudhoe JE, Jorgensen B, Ihrke G, Doyonnas R, Zannettino AC, Buckle VJ, Ward CJ, Simmons PJ, Watt SM
JournalJ Biol Chem
PubMed ID11027692
'Functional analyses have indicated that the human CD164 sialomucin may play a key role in hematopoiesis by facilitating the adhesion of human CD34(+) cells to the stroma and by negatively regulating CD34(+)CD38(lo/-) cell proliferation. We have identified three novel human CD164 variants derived by alternative splicing of bona fide exons ... More
Involvement of phosphorylation in doxorubicin-mediated myofibril degeneration. An immunofluorescence microscopy analysis.
AuthorsSussman MA, Hamm-Alvarez SF, Vilalta PM, Welch S, Kedes L
JournalCirc Res
PubMed ID8978322
'Loss of myofilaments has been observed in both adaptive cardiac responses (i.e., hypertrophy) as well as in chemotheraputic use of antineoplastic drugs with cardiotoxic side effects (i.e., doxorubicin). An understanding of the degenerative process is a prerequisite for determining approaches to limit the cardiomyopathic changes associated with chronic heart disease ... More
Filamin 2 (FLN2): A muscle-specific sarcoglycan interacting protein.
AuthorsThompson TG, Chan YM, Hack AA, Brosius M, Rajala M, Lidov HG, McNally EM, Watkins S, Kunkel LM
JournalJ Cell Biol
PubMed ID10629222
'Mutations in genes encoding for the sarcoglycans, a subset of proteins within the dystrophin-glycoprotein complex, produce a limb-girdle muscular dystrophy phenotype; however, the precise role of this group of proteins in the skeletal muscle is not known. To understand the role of the sarcoglycan complex, we looked for sarcoglycan interacting ... More
Tissue transglutaminase mediates activation of RhoA and MAP kinase pathways during retinoic acid-induced neuronal differentiation of SH-SY5Y cells.
AuthorsSingh US, Pan J, Kao YL, Joshi S, Young KL, Baker KM
JournalJ Biol Chem
PubMed ID12401808
'All-trans-retinoic acid (RA) plays a crucial role in survival and differentiation of neurons. For elucidating signaling mechanisms involved in RA-induced neuronal differentiation, we have selected SH-SY5Y cells, which are an established in vitro cell model for studying RA signaling. Here we report that RA-induced neuronal differentiation of SH-SY5Y cells is ... More
Plasma membrane Ca2+-atpase isoforms 2b and 4b interact promiscuously and selectively with members of the membrane-associated guanylate kinase family of PDZ (PSD95/Dlg/ZO-1) domain-containing proteins.
AuthorsDeMarco SJ, Strehler EE
JournalJ Biol Chem
PubMed ID11274188
'Spatial and temporal regulation of intracellular Ca(2+) signaling depends on localized Ca(2+) microdomains containing the requisite molecular components for Ca(2+) influx, efflux, and signal transmission. Plasma membrane Ca(2+)-ATPase (PMCA) isoforms of the "b" splice type contain predicted PDZ (PSD95/Dlg/ZO-1) interaction domains. The COOH-terminal tail of PMCA2b isolated the membrane-associated guanylate ... More
Mass spectrometric analysis of GAP-43/neuromodulin reveals the presence of a variety of fatty acylated species.
Authors Liang Xiquan; Lu Yun; Neubert Thomas A; Resh Marilyn D;
JournalJ Biol Chem
PubMed ID12105219
'GAP-43 (neuromodulin) is a protein kinase C substrate that is abundant in developing and regenerating neurons. Thioester-linked palmitoylation at two cysteines near the GAP-43 N terminus has been implicated in directing membrane binding. Here, we use mass spectrometry to examine the stoichiometry of palmitoylation and the molecular identity of the ... More
Galpha 13 requires palmitoylation for plasma membrane localization, Rho-dependent signaling, and promotion of p115-RhoGEF membrane binding.
AuthorsBhattacharyya R, Wedegaertner PB
JournalJ Biol Chem
PubMed ID10747909
'Most heterotrimeric G protein alpha subunits are covalently modified by palmitate attached to one or more N-terminal cysteine residues. Although a wide variety of proteins undergo palmitoylation, the role of this fatty acid modification in G protein signaling is not well understood. Thus, we examined the role of palmitoylation of ... More
Calcyon, a novel partner of clathrin light chain, stimulates clathrin-mediated endocytosis.
AuthorsXiao J, Dai R, Negyessy L, Bergson C
JournalJ Biol Chem
PubMed ID16595675
'In the central nervous system, clathrin-mediated endocytosis is crucial for efficient synaptic transmission. Clathrin-coated vesicle assembly and disassembly is regulated by some 30 adaptor and accessory proteins, most of which interact with clathrin heavy chain. Using the calcyon cytosolic domain as bait, we isolated clathrin light chain in a yeast ... More
The ligand for osteoprotegerin (OPGL) directly activates mature osteoclasts.
AuthorsBurgess TL, Qian Y, Kaufman S, Ring BD, Van G, Capparelli C, Kelley M, Hsu H, Boyle WJ, Dunstan CR, Hu S, Lacey DL
JournalJ Cell Biol
PubMed ID10225954
'Osteoprotegerin (OPG) and OPG-ligand (OPGL) potently inhibit and stimulate, respectively, osteoclast differentiation (Simonet, W.S., D.L. Lacey, C.R. Dunstan, M. Kelley, M.-S. Chang, R. Luethy, H.Q. Nguyen, S. Wooden, L. Bennett, T. Boone, et al. 1997. Cell. 89:309-319; Lacey, D.L., E. Timms, H.-L. Tan, M.J. Kelley, C.R. Dunstan, T. Burgess, R. ... More
A PMLRARalpha transgene initiates murine acute promyelocytic leukemia.
AuthorsBrown D, Kogan S, Lagasse E, Weissman I, Alcalay M, Pelicci PG, Atwater S, Bishop JM
JournalProc Natl Acad Sci U S A
PubMed ID9122233
'The malignant cells of acute promyelocytic leukemia (APL) contain a reciprocal chromosomal translocation that fuses the promyelocytic leukemia gene (PML) with the retinoic acid receptor alpha gene (RAR alpha). To test the hypothesis that the chimera PMLRAR alpha plays a role in leukemogenesis, we expressed a PMLRAR alpha cDNA in ... More
A dynactin subunit with a highly conserved cysteine-rich motif interacts directly with Arp1.
AuthorsKarki S, Tokito MK, Holzbaur EL
JournalJ Biol Chem
PubMed ID10671518
'Dynactin is a multisubunit complex and a required cofactor for most, or all, of the cellular processes powered by the microtubule-based motor cytoplasmic dynein. Using a dynein affinity column, the previously uncharacterized p62 subunit of dynactin was isolated and microsequenced. Two peptide sequences were used to clone human cDNAs encoding ... More
Deletion analogues of transportan.
AuthorsSoomets U, Lindgren M, Gallet X, Hällbrink M, Elmquist A, Balaspiri L, Zorko M, Pooga M, Brasseur R, Langel U
JournalBiochim Biophys Acta
PubMed ID10930519
'Several shorter analogues of the cell penetrating peptide, transportan, have been synthesized in order to define the regions of the sequence, which are responsible for the membrane translocation property of the peptide. Penetration of the peptides into Bowes melanoma cells and the influence on GTPase activity in Rin m5F cellular ... More
Comparison of methods for erythroblast selection: application to selecting fetal erythroblasts from maternal blood.
AuthorsCollarini EJ, Cain CA, Gammon D, Harriman B, Magee K, Du G, Schueler PA, Mahoney WC
JournalCytometry
PubMed ID11746096
'BACKGROUND: Many methods have been employed to obtain fetal cells from maternal blood for prenatal diagnostics, but there has been little work done that compares the efficacy of different methods. This study presents a comparison of two commonly used methods for selecting erythroblasts with selection directly from whole blood. METHODS: ... More
Critical role of serine 465 in isoflurane-induced increase of cell-surface redistribution and activity of glutamate transporter type 3.
AuthorsHuang Y, Feng X, Sando JJ, Zuo Z
JournalJ Biol Chem
PubMed ID17062570
'Glutamate transporters (also called excitatory amino acid transporters, EAATs) bind extracellular glutamate and transport it to intracellular space to regulate glutamate neurotransmission and to maintain extracellular glutamate concentrations below neurotoxic levels. We previously showed that isoflurane, a commonly used anesthetic, enhanced the activity of EAAT3, a major neuronal EAAT. This ... More
Reaction diffusion modeling of calcium dynamics with realistic ER geometry.
AuthorsMeans S, Smith AJ, Shepherd J, Shadid J, Fowler J, Wojcikiewicz RJ, Mazel T, Smith GD, Wilson BS
JournalBiophys J
PubMed ID16617072
'We describe a finite-element model of mast cell calcium dynamics that incorporates the endoplasmic reticulum''s complex geometry. The model is built upon a three-dimensional reconstruction of the endoplasmic reticulum (ER) from an electron tomographic tilt series. Tetrahedral meshes provide volumetric representations of the ER lumen, ER membrane, cytoplasm, and plasma ... More
Distribution of CTP:phosphocholine cytidylyltransferase (CCT) isoforms. Identification of a new CCTbeta splice variant.
AuthorsLykidis A, Baburina I, Jackowski S
JournalJ Biol Chem
PubMed ID10480912
'CTP:phosphocholine cytidylyltransferase is a major regulator of phosphatidylcholine biosynthesis. A single isoform, CCTalpha, has been studied extensively and a second isoform, CCTbeta, was recently identified. We identify and characterize a third cDNA, CCTbeta2, that differs from CCTbeta1 at the carboxyl-terminal end and is predicted to arise as a splice variant ... More
Regulated trafficking of the human dopamine transporter. Clathrin-mediated internalization and lysosomal degradation in response to phorbol esters.
AuthorsDaniels GM, Amara SG
JournalJ Biol Chem
PubMed ID10585462
'The dopamine transporter plays an essential role in the modulation of dopaminergic neurotransmission by mediating the reuptake of dopamine into presynaptic neurons. In cells expressing the dopamine transporter, activation of protein kinase C by phorbol esters results in a significant reduction in dopamine uptake. This phorbol ester-mediated inhibition of dopamine ... More
Philinopside E, a new sulfated saponin from sea cucumber, blocks the interaction between kinase insert domain-containing receptor (KDR) and alphavbeta3 integrin via binding to the extracellular domain of KDR.
AuthorsTian F, Zhu CH, Zhang XW, Xie X, Xin XL, Yi YH, Lin LP, Geng MY, Ding J,
JournalMol Pharmacol
PubMed ID17565003
'Vascular endothelial growth factor (VEGF) signaling pathway is essential for tumor angiogenesis and has long been recognized as a promising target for cancer therapy. Current view holds that physical interaction between alpha(v)beta(3) integrin and kinase insert domain-containing receptor (KDR) is important in regulating angiogenesis and tumor development. We have reported ... More
Detection and analysis of tumor fluorescence using a two-photon optical fiber probe.
AuthorsThomas TP, Myaing MT, Ye JY, Candido K, Kotlyar A, Beals J, Cao P, Keszler B, Patri AK, Norris TB, Baker JR
JournalBiophys J
PubMed ID15189892
'The utility of a two-photon optical fiber fluorescence probe (TPOFF) for sensing and quantifying tumor fluorescent signals was tested in vivo. Xenograft tumors were developed in athymic mice using MCA207 cells expressing green fluorescent protein (GFP). The TPOFF probe was able to detect ex vivo fluorescence from excised tumors containing ... More
Multiphoton excitation spectra in biological samples.
AuthorsDickinson ME, Simbuerger E, Zimmermann B, Waters CW, Fraser SE
JournalJ Biomed Opt
PubMed ID12880336
'Multiphoton microscopy is becoming a popular mode of live and fixed cell imaging. This mode of imaging offers several advantages due to the fact that fluorochrome excitation is a nonlinear event resulting in excitation only at the plane of focus. Multiphoton excitation is enhanced by the use of ultrafast lasers ... More
Bcl-x(S) anatagonizes the protective effects of Bcl-x(L).
AuthorsMinn AJ, Boise LH, Thompson CB
JournalJ Biol Chem
PubMed ID8626425
'Bcl-x, a member of the Bcl-2 family, has two alternatively spliced forms, Bcl-x(L) and Bcl-x(S). Bcl-x(L), like Bcl-2, is able to protect cells from a wide variety of apoptotic stimuli. Bcl-x(S), as a result of alternative splicing, lacks 63 amino acids that comprise the region of greatest amino acid identity ... More
An anti-transferrin receptor-avidin fusion protein exhibits both strong proapoptotic activity and the ability to deliver various molecules into cancer cells.
AuthorsNg PP, Dela Cruz JS, Sorour DN, Stinebaugh JM, Shin SU, Shin DS, Morrison SL, Penichet ML
JournalProc Natl Acad Sci U S A
PubMed ID12149472
'We have developed an antibody fusion protein (anti-rat TfR IgG3-Av) with the ability to deliver different molecules into cancer cells. It consists of avidin genetically fused to the C(H)3 region of a human IgG3 specific for the rat transferrin receptor. It forms strong, noncovalent interactions with biotinylated molecules such as ... More
Activities of the microtubule-stabilizing agents epothilones A and B with purified tubulin and in cells resistant to paclitaxel (Taxol(R)).
AuthorsKowalski RJ, Giannakakou P, Hamel E
JournalJ Biol Chem
PubMed ID8999970
'Epothilones A and B, natural products with minimal structural analogy to taxoids, have effects similar to those of paclitaxel (Taxol(R)) in cultured cells and on microtubule protein, but differ from paclitaxel in retaining activity in multidrug-resistant cells. We examined interactions of the epothilones with purified tubulin and additional cell lines, ... More
Sensory neurons and schwann cells respond to oxidative stress by increasing antioxidant defense mechanisms.
AuthorsVincent AM, Kato K, McLean LL, Soules ME, Feldman EL,
JournalAntioxid Redox Signal
PubMed ID19072199
'Elevated blood glucose is a key initiator of mechanisms leading to diabetic neuropathy. Increases in glucose induce acute mitochondrial oxidative stress in dorsal root ganglion (DRG) neurons, the sensory neurons normally affected in diabetic neuropathy, whereas Schwann cells are largely unaffected. We propose that activation of an antioxidant response in ... More
Inducible expression of a dominant negative DNA polymerase-gamma depletes mitochondrial DNA and produces a rho0 phenotype.
AuthorsJazayeri M, Andreyev A, Will Y, Ward M, Anderson CM, Clevenger W,
JournalJ Biol Chem
PubMed ID12645575
'We report the inducible, stable expression of a dominant negative form of mitochondria-specific DNA polymerase-gamma to eliminate mitochondrial DNA (mtDNA) from human cells in culture. HEK293 cells were transfected with a plasmid encoding inactive DNA polymerase-gamma harboring a D1135A substitution (POLGdn). The cells rapidly lost mtDNA (t1/2 = 2-3 days) ... More
Quality assessment of confocal microscopy slide based systems: performance.
AuthorsZucker RM
JournalCytometry A
PubMed ID16807897
'BACKGROUND: All fluorescence slide-based cytometry detections systems basically include the following components: (1) an excitation light source, (2) intermediate optics, and (3) a detection device consisting of a CCD camera or a PMT. The optical principles employed is slide-based systems are similar to those of confocal microscopes (CLSM). METHODS: The ... More
The effects of osmotic stress on the viscoelastic and physical properties of articular chondrocytes.
AuthorsGuilak F, Erickson GR, Ting-Beall HP
JournalBiophys J
PubMed ID11806914
'The metabolic activity of chondrocytes in articular cartilage is influenced by alterations in the osmotic environment of the tissue, which occur secondary to mechanical compression. The mechanism by which osmotic stress modulates cell physiology is not fully understood and may involve changes in the physical properties of the membrane or ... More
Measurement of DNA damage associated with apoptosis by laser scanning cytometry.
AuthorsBacso Z, Eliason JF
JournalCytometry
PubMed ID11746086
'BACKGROUND: Phosphatidylserine (PS) binding by annexin V (AV) is an early membrane marker of apoptosis. Using laser scanning cytometry (LSC) and the comet assay, we showed that the DNA of AV(+) cells is so highly fragmented that it cannot be quantified by the comet assay (Bacso et al.: Cancer Res ... More
IL-6 produced by macrophages infected with Mycobacterium species suppresses T cell responses.
AuthorsVanHeyningen TK, Collins HL, Russell DG
JournalJ Immunol
PubMed ID8977207
'The ability of Mycobacterium bovis Calmette-Guérin bacillus-infected bone marrow-derived macrophages to process and present exogenously added Ags to T cells and stimulate their growth and production of IL-2 was examined. The infected macrophages were inhibited in their ability to activate T cells, and this inhibition could be transferred to uninfected ... More
Synthesis and identification of small molecules that potently induce apoptosis in melanoma cells through G1 cell cycle arrest.
'Late-stage malignant melanoma is a cancer that is refractory to current chemotherapeutic treatments. The average survival time for patients with such a diagnosis is 6 months. In general, the vast majority of anticancer drugs operate through induction of cell cycle arrest and cell death in either the DNA synthesis (S) ... More
Assessment of telomere length in hematopoietic interphase cells using in situ hybridization and digital fluorescence microscopy.
'Telomeres are G/C-rich repetitive DNA sequences at the end of all eukaryotic chromosomes. The loss of telomeric repeat sequences during cell divisions has been proposed as a possible mechanism for cell senescence. The standard procedure for measurement of telomere length is Southern blot (SB) hybridization with a telomere-specific probe. However, ... More
Telomere length assessment in human archival tissues: combined telomere fluorescence in situ hybridization and immunostaining.
AuthorsMeeker AK, Gage WR, Hicks JL, Simon I, Coffman JR, Platz EA, March GE, De Marzo AM
JournalAm J Pathol
PubMed ID11943711
'A method was developed to assess human telomere lengths at the individual cell level in tissue sections from standard formalin-fixed paraffin-embedded tissues. We coupled this method with immunofluorescence to allow the simultaneous identification of specific cell types. Validation of this in situ quantification method showed excellent agreement with the commonly ... More
Antifading agents for confocal fluorescence microscopy.
AuthorsBerrios M, Conlon KA, Colflesh DE
JournalMethods Enzymol
PubMed ID10506967
Green fluorescent protein (GFP): applications in cell-based assays for drug discovery.
AuthorsKain SR
JournalDrug Discov Today
PubMed ID10377508
Green fluorescent protein (GFP) is a powerful tool for cell-based assays owing to the intrinsic fluorescence of this protein that allows real-time analysis of molecular events in living cells. A number of GFP variants have been developed with optimal properties for both high-throughput screening and high-content screening. The author discusses ... More
Multiplex detection of RNA expression in Drosophila embryos.
AuthorsKosman D, Mizutani CM, Lemons D, Cox WG, McGinnis W, Bier E
JournalScience
PubMed ID15297669
We present a fluorescence-based, multiplex in situ hybridization method that permits the simultaneous detection of five differently labeled antisense RNA probes and up to seven differ-ent transcripts in a single Drosophila embryo. We also show that it should be possible to increase the number of detected transcripts substantially with nascent ... More
A pool of extramitochondrial frataxin that promotes cell survival.
AuthorsCondò I, Ventura N, Malisan F, Tomassini B, Testi R
JournalJ Biol Chem
PubMed ID16608849
Frataxin is a mitochondrial protein involved in iron metabolism. Defective expression of frataxin causes Friedreich ataxia (FA), an inherited degenerative syndrome characterized by ataxia, cardiomyopathy, and high incidence of diabetes. Here we report that frataxin-deficient cells are more prone to undergo stress-induced mitochondrial damage and apoptosis, while the overexpression of ... More
Hyaluronic acid-N-hydroxysuccinimide: a useful intermediate for bioconjugation.
AuthorsLuo Y, Prestwich GD
JournalBioconjug Chem
PubMed ID11716704
Hyaluronic acid (HA) is an abundant nonsulfated glycosaminoglycan component of synovial fluid and the extracellular matrix. HA is an important building block for biocompatible and biointeractive materials with applications in drug delivery, tissue engineering, wound repair, and viscosupplementation. Herein we describe the synthesis and characterization of HA-N-succinimide, an activated ester ... More
In vivo models for studying mast cell-dependent responses to bacterial infection.
AuthorsShelburne CP, McLachlan JB, Abraham SN,
JournalMethods Mol Biol
PubMed ID16110170
Mast cells are a critical component of host defense against bacterial infections. Activation of these cells during infection induces both innate and adaptive aspects of protective immunity needed for the elimination of the bacteria and survival of the host. These functional roles for the mast cell have been principally characterized ... More
Argosomes: a potential vehicle for the spread of morphogens through epithelia.
AuthorsGreco V, Hannus M, Eaton S
JournalCell
PubMed ID11551510
The formation of morphogen gradients is essential for tissue patterning. Morphogens are released from producing cells and spread through adjacent tissue; paradoxically, however, many morphogens, including Wingless, associate tightly with the cell membrane. Here, we describe a novel cell biological mechanism that disperses membrane fragments over large distances through the ... More
Direct eye visualization of Cy5 fluorescence for immunocytochemistry and in situ hybridization.
AuthorsFerri GL, Isola J, Berger P, Giro G
JournalJ Histochem Cytochem
PubMed ID10681398
Cyanine 5.18 (or Cy5) is a fluorochrome emitting in the long-red/far-red range, usually regarded as unsuitable for direct observation by the human eye. We describe here the optimization of a direct visualization approach to Cy5 labeling, based on a standard fluorescence microscope with mercury light excitation and applicable to both ... More
The identification and characterization of two phosphatidylinositol-4,5-bisphosphate 4-phosphatases.
AuthorsUngewickell A, Hugge C, Kisseleva M, Chang SC, Zou J, Feng Y, Galyov EE, Wilson M, Majerus PW
JournalProc Natl Acad Sci U S A
PubMed ID16365287
Numerous inositol polyphosphate 5-phosphatases catalyze the degradation of phosphatidylinositol-4,5-bisphosphate (PtdIns-4,5-P(2)) to phosphatidylinositol-4-phosphate (PtdIns-4-P). An alternative pathway to degrade PtdIns-4,5-P(2) is the hydrolysis of PtdIns-4,5-P(2) by a 4-phosphatase, leading to the production of PtdIns-5-P. Whereas the bacterial IpgD enzyme is known to catalyze this reaction, no such mammalian enzyme has been ... More
CD19 targeting of chronic lymphocytic leukemia with a novel Fc-domain-engineered monoclonal antibody.
AuthorsAwan FT, Lapalombella R, Trotta R, Butchar JP, Yu B, Benson DM, Roda JM, Cheney C, Mo X, Lehman A, Jones J, Flynn J, Jarjoura D, Desjarlais JR, Tridandapani S, Caligiuri MA, Muthusamy N, Byrd JC,
JournalBlood
PubMed ID19965644
CD19 is a B cell-specific antigen expressed on chronic lymphocytic leukemia (CLL) cells but to date has not been effectively targeted with therapeutic monoclonal antibodies. XmAb5574 is a novel engineered anti-CD19 monoclonal antibody with a modified constant fragment (Fc)-domain designed to enhance binding of FcgammaRIIIa. Herein, we demonstrate that XmAb5574 ... More
Triton X-100 pretreatment of LR-white thin sections improves immunofluorescence specificity and intensity.
AuthorsGhrebi SS, Owen GR, Brunette DM,
JournalMicrosc Res Tech
PubMed ID17380496
The staining of intracellular antigenic sites in postembedded samples is a challenging problem. Deterioration of antigenicity and limited antibody accessibility to the antigen are commonly encountered on account of processing steps. In this study preservation of the antigen was achieved by fixing the tissues with mild fixatives, performing partial dehydration, ... More
Altered lipid droplet dynamics in hepatocytes lacking triacylglycerol hydrolase expression.
AuthorsWang H, Wei E, Quiroga AD, Sun X, Touret N, Lehner R,
JournalMol Biol Cell
PubMed ID20410140
Lipid droplets (LDs) form from the endoplasmic reticulum (ER) and grow in size by obtaining triacylglycerols (TG). Triacylglycerol hydrolase (TGH), a lipase residing in the ER, is involved in the mobilization of TG stored in LDs for the secretion of very-low-density lipoproteins. In this study, we investigated TGH-mediated changes in ... More
Apoptosis induces efflux of the mitochondrial matrix enzyme deoxyguanosine kinase.
AuthorsJüllig M, Eriksson S,
JournalJ Biol Chem
PubMed ID11294860
Deoxyguanosine kinase (dGK) initiates the salvage of purine deoxynucleosides in mitochondria and is a key enzyme in mitochondrial DNA precursor synthesis. The active form of the enzyme is a 60-kDa protein normally located in the mitochondrial matrix. Here we describe the subcellular distribution of dGK during apoptosis in human epithelial ... More
Cytogenetic evidence for asexual evolution of bdelloid rotifers.
AuthorsMark Welch JL, Mark Welch DB, Meselson M
JournalProc Natl Acad Sci U S A
PubMed ID14747655
DNA sequencing has shown individual bdelloid rotifer genomes to contain two or more diverged copies of every gene examined and has revealed no closely similar copies. These and other findings are consistent with long-term asexual evolution of bdelloids. It is not entirely ruled out, however, that bdelloid genomes consist of ... More
The origin recognition core complex regulates dendrite and spine development in postmitotic neurons.
AuthorsHuang Z, Zang K, Reichardt LF
JournalJ Cell Biol
PubMed ID16087709
The origin recognition complex (ORC) ensures exactly one round of genome replication per cell cycle through acting as a molecular switch that precisely controls the assembly, firing, and inactivation of the replication initiation machinery. Recent data indicate that it may also coordinate the processes of mitosis and cytokinesis and ensure ... More
Ultrathin cryosections: an important tool for immunofluorescence and correlative microscopy.
AuthorsTakizawa T, Robinson JM
JournalJ Histochem Cytochem
PubMed ID12754282
Here we show that ultrathin cryosections of placental tissue can be used as a substrate in immunofluorescence experiments. A high degree of spatial resolution can be achieved in these preparations because there is essentially no out-of-focus fluorescence. Therefore, immunofluorescence microscopy using ultrathin cryosections provides a very useful method for determining ... More
Laminin-5 induces osteogenic gene expression in human mesenchymal stem cells through an ERK-dependent pathway.
AuthorsKlees RF, Salasznyk RM, Kingsley K, Williams WA, Boskey A, Plopper GE
JournalMol Biol Cell
PubMed ID15574877
The laminin family of proteins is critical for managing a variety of cellular activities including migration, adhesion, and differentiation. In bone, the roles of laminins in controlling osteogenic differentiation of human mesenchymal stem cells (hMSC) are unknown. We report here that laminin-5 is found in bone and expressed by hMSC. ... More
Validation and quantitation of an in vitro M-cell model.
AuthorsTyrer P, Ruth Foxwell A, Kyd J, Harvey M, Sizer P, Cripps A
JournalBiochem Biophys Res Commun
PubMed ID12445810
This study has evaluated an in vitro model of the follicle-associated epithelia that overlie Peyer's patches of the small intestine. The model shares many phenotypic characteristics of M cells in vivo. Co-cultures of the human adenocarcinoma cell line Caco-2 and freshly isolated Peyer's patch cells were established. Fluorescence microscopy and ... More
The Enterococcus faecalis fsr two-component system controls biofilm development through production of gelatinase.
AuthorsHancock LE, Perego M
JournalJ Bacteriol
PubMed ID15317767
Bacterial growth as a biofilm on solid surfaces is strongly associated with the development of human infections. Biofilms on native heart valves (infective endocarditis) is a life-threatening disease as a consequence of bacterial resistance to antimicrobials in such a state. Enterococci have emerged as a cause of endocarditis and nosocomial ... More
Direct extracellular contact between integrin alpha(3)beta(1) and TM4SF protein CD151.
AuthorsYauch RL, Kazarov AR, Desai B, Lee RT, Hemler ME
JournalJ Biol Chem
PubMed ID10734060
Previously we established that the alpha(3)beta(1) integrin shows stable, specific, and stoichiometric association with the TM4SF (tetraspannin) protein CD151. Here we used a membrane impermeable cross-linking agent to show a direct association between extracellular domains of alpha(3)beta(1) and CD151. The alpha(3)beta(1)-CD151 association site was then mapped using chimeric alpha(6)/alpha(3) integrins ... More
Bcl-2 inhibits a Fas-induced conformational change in the Bax N terminus and Bax mitochondrial translocation.
AuthorsMurphy KM, Streips UN, Lock RB
JournalJ Biol Chem
PubMed ID10751382
Members of the Bcl-2 family of proteins control the cellular commitment to apoptosis, although their role in Fas-induced apoptosis is ill-defined. In this report we demonstrate that activation of the Fas receptor present on a human breast epithelial cell line resulted in a conformational change in the N terminus of ... More
Phosphate deprivation induces transfer of DGDG galactolipid from chloroplast to mitochondria.
AuthorsJouhet J, Maréchal E, Baldan B, Bligny R, Joyard J, Block MA
JournalJ Cell Biol
PubMed ID15569715
In many soils plants have to grow in a shortage of phosphate, leading to development of phosphate-saving mechanisms. At the cellular level, these mechanisms include conversion of phospholipids into glycolipids, mainly digalactosyldiacylglycerol (DGDG). The lipid changes are not restricted to plastid membranes where DGDG is synthesized and resides under normal ... More
Immobilization of bioactive fibroblast growth factor-2 into cubic proteinous microcrystals (Bombyx mori cypovirus polyhedra) that are insoluble in a physiological cellular environment.
AuthorsMori H, Shukunami C, Furuyama A, Notsu H, Nishizaki Y, Hiraki Y
JournalJ Biol Chem
PubMed ID17430896
The supramolecular architecture of the extracellular matrix and the disposition of its specific accessory molecules give rise to variable heterotopic signaling cues for single cells. Here we have described the successful occlusion of human fibroblast growth factor-2 (FGF-2) into the cubic inclusion bodies (FGF-2 polyhedra) of the Bombyx mori cytoplasmic ... More
TAP-independent presentation of CTL epitopes by Trojan antigens.
AuthorsLu J, Wettstein PJ, Higashimoto Y, Appella E, Celis E
JournalJ Immunol
PubMed ID11390450
The majority of CTL epitopes are derived from intracellular proteins that are degraded in the cytoplasm by proteasomes into peptides that are transported into the endoplasmic reticulum by the TAP complex. These peptides can be further processed into the optimal size (8-10 residues) for binding with nascent MHC class I ... More
The enzyme 12/15-lipoxygenase (12/15-LO) introduces peroxyl groups in a position-specific manner into unsaturated fatty acids in certain cells, but the role of such enzymatic lipid peroxidation remains poorly defined. Here we report a novel function for 12/15-LO in mouse peritoneal macrophages. When macrophages were coincubated with apoptotic cells, the enzyme ... More
Posttranslational myristoylation of caspase-activated p21-activated protein kinase 2 (PAK2) potentiates late apoptotic events.
AuthorsVilas GL, Corvi MM, Plummer GJ, Seime AM, Lambkin GR, Berthiaume LG
JournalProc Natl Acad Sci U S A
PubMed ID16617111
p21-activated protein kinase (PAK) 2 is a small GTPase-activated serine/threonine kinase regulating various cytoskeletal functions and is cleaved by caspase-3 during apoptosis. We demonstrate that the caspase-cleaved PAK2 C-terminal kinase fragment (C-t-PAK2) is posttranslationally myristoylated, although myristoylation is typically a cotranslational process. Myristoylation and an adjacent polybasic domain of C-t-PAK2 ... More
ErbB-2 amplification inhibits down-regulation and induces constitutive activation of both ErbB-2 and epidermal growth factor receptors.
AuthorsWorthylake R, Opresko LK, Wiley HS
JournalJ Biol Chem
PubMed ID10085130
ErbB-2/HER2 is an important signaling partner for the epidermal growth factor receptor (EGFR). Overexpression of erbB-2 is also associated with poor prognosis in breast cancer. To investigate how erbB-2 amplification affects its interactions with the EGFR, we used a human mammary epithelial cell system in which erbB-2 expression was increased ... More
Mammalian p55CDC mediates association of the spindle checkpoint protein Mad2 with the cyclosome/anaphase-promoting complex, and is involved in regulating anaphase onset and late mitotic events.
AuthorsKallio M, Weinstein J, Daum JR, Burke DJ, Gorbsky GJ
JournalJ Cell Biol
PubMed ID9628895
We have investigated the function of p55CDC, a mammalian protein related to Cdc20 and Hct1/Cdh1 in Saccharomyces cerevisiae, and Fizzy and Fizzy-related in Drosophila. Immunofluorescence studies and expression of a p55CDC-GFP chimera demonstrate that p55CDC is concentrated at the kinetochores in M phase cells from late prophase to telophase. Some ... More
Calcium influx through AMPA receptors and through calcium channels is regulated by protein kinase C in cultured retina amacrine-like cells.
AuthorsCarvalho AL, Duarte CB, Faro CJ, Carvalho AP, Pires EV
JournalJ Neurochem
PubMed ID9572298
The functional modulation of the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptors by protein kinase C (PKC) was investigated in cultures enriched in retinal amacrine-like cells. The kainate-evoked [Ca2+]i increase is due to Ca2+ entry through open AMPA receptor channels, because it was blocked by the active isomer of a 2,3-benzodiazepine (LY 303070), an ... More
Prenylated Rab acceptor protein is a receptor for prenylated small GTPases.
AuthorsFigueroa C, Taylor J, Vojtek AB
JournalJ Biol Chem
PubMed ID11335720
Localization of Ras and Ras-like proteins to the correct subcellular compartment is essential for these proteins to mediate their biological effects. Many members of the Ras superfamily (Ha-Ras, N-Ras, TC21, and RhoA) are prenylated in the cytoplasm and then transit through the endomembrane system on their way to the plasma ... More
Seizures accelerate functional integration of adult-generated granule cells.
AuthorsOverstreet-Wadiche LS, Bromberg DA, Bensen AL, Westbrook GL
JournalJ Neurosci
PubMed ID16611826
In humans and experimental animals, structural and functional changes in neural circuits can accompany the development of epilepsy. In the dentate gyrus, seizures enhance adult neurogenesis, but it is unclear to what extent newborn granule cells participate in seizure-induced synaptic reorganization. During the first weeks of their existence, mouse newborn ... More
A novel specific role for I kappa B kinase complex-associated protein in cytosolic stress signaling.
We demonstrate here a novel role for the I kappa B kinase complex-associated protein (IKAP) in the regulation of activation of the mammalian stress response via the c-Jun N-terminal kinase (JNK)-signaling pathway. We cloned IKAP as a JNK-associating protein using the Ras recruitment yeast two-hybrid system. IKAP efficiently and specifically ... More
ATPase-defective mammalian VPS4 localizes to aberrant endosomes and impairs cholesterol trafficking.
AuthorsBishop N, Woodman P
JournalMol Biol Cell
PubMed ID10637304
The yeast vacuolar sorting protein Vps4p is an ATPase required for endosomal trafficking that couples membrane association to its ATPase cycle. To investigate the function of mammalian VPS4 in endosomal trafficking, we have transiently expressed wild-type or ATPase-defective human VPS4 (hVPS4) in cultured cells. Wild-type hVPS4 was cytosolic, whereas a ... More
Release of kinesin from vesicles by hsc70 and regulation of fast axonal transport.
AuthorsTsai MY, Morfini G, Szebenyi G, Brady ST
JournalMol Biol Cell
PubMed ID10848636
The nature of kinesin interactions with membrane-bound organelles and mechanisms for regulation of kinesin-based motility have both been surprisingly difficult to define. Most kinesin is recovered in supernatants with standard protocols for purification of motor proteins, but kinesin recovered on membrane-bound organelles is tightly bound. Partitioning of kinesin between vesicle ... More
Two ZBP1 KH domains facilitate beta-actin mRNA localization, granule formation, and cytoskeletal attachment.
Chicken embryo fibroblasts (CEFs) localize beta-actin mRNA to their lamellae, a process important for the maintenance of cell polarity and motility. The localization of beta-actin mRNA requires a cis localization element (zipcode) and involves zipcode binding protein 1 (ZBP1), a protein that specifically binds to the zipcode. Both localize to ... More
rab7 activity affects epidermal growth factor:epidermal growth factor receptor degradation by regulating endocytic trafficking from the late endosome.
AuthorsCeresa BP, Bahr SJ
JournalJ Biol Chem
PubMed ID16282324
The epidermal growth factor receptor (EGFR) is a member of the receptor tyrosine kinase family. Ligand (epidermal growth factor or EGF) binding to the EGFR results in the coordinated activation and integration of biochemical signaling events to mediate cell growth, migration, and differentiation. One mechanism the cell utilizes to orchestrate ... More
MHC class II-peptide complexes and APC lipid rafts accumulate at the immunological synapse.
AuthorsHiltbold EM, Poloso NJ, Roche PA
JournalJ Immunol
PubMed ID12538693
Activation of CD4(+) Th cells requires their cognate interaction with APCs bearing specific relevant MHC class II-peptide complexes. This cognate interaction culminates in the formation of an immunological synapse that contains the various proteins and lipids required for efficient T cell activation. We now show that APC lipid raft membrane ... More
Extracellular application of nicotinic acid adenine dinucleotide phosphate induces Ca2+ signaling in astrocytes in situ.
AuthorsHeidemann AC, Schipke CG, Kettenmann H
JournalJ Biol Chem
PubMed ID16061474
Nicotinic acid adenine dinucleotide phosphate (NAADP+) has been identified as a novel second messenger triggering Ca2+ release from intracellular stores. Here we report that murine cortical astrocytes in culture and in acute slices respond with transient intracellular Ca2+ increases to extracellularly applied NAADP+ and express the NAADP+-producing enzyme CD38. The ... More
c-erbB-3: a nuclear protein in mammary epithelial cells.
AuthorsOffterdinger M, Schöfer C, Weipoltshammer K, Grunt TW
JournalJ Cell Biol
PubMed ID12045181
c-erbB receptors are usually located in cell membranes and are activated by extracellular binding of EGF-like growth factors. Unexpectedly, using immunofluorescence we found high levels of c-erbB-3 within the nuclei of MTSV1-7 immortalized nonmalignant human mammary epithelial cells. Nuclear localization was mediated by the COOH terminus of c-erbB-3, and a ... More
Rapid analysis of mitochondrial DNA depletion by fluorescence in situ hybridization and immunocytochemistry: potential strategies for HIV therapeutic monitoring.
AuthorsJanes MS, Hanson BJ, Hill DM, Buller GM, Agnew JY, Sherwood SW, Cox WG, Yamagata K, Capaldi RA
JournalJ Histochem Cytochem
PubMed ID15258176
Nucleoside reverse transcriptase inhibitors (NRTIs) have been a mainstay in the treatment of human immunodeficiency virus since the introduction of azidothymidine (AZT) in 1987. However, none of the current therapies can completely eradicate the virus, necessitating long-term use of anti-retroviral drugs to prevent viral re-growth. One of the side effects ... More
Distinct membrane domains on endosomes in the recycling pathway visualized by multicolor imaging of Rab4, Rab5, and Rab11.
AuthorsSönnichsen B, De Renzis S, Nielsen E, Rietdorf J, Zerial M
JournalJ Cell Biol
PubMed ID10811830
Two endosome populations involved in recycling of membranes and receptors to the plasma membrane have been described, the early and the recycling endosome. However, this distinction is mainly based on the flow of cargo molecules and the spatial distribution of these membranes within the cell. To get insights into the ... More
Deafness and stria vascularis defects in S1P2 receptor-null mice.
AuthorsKono M, Belyantseva IA, Skoura A, Frolenkov GI, Starost MF, Dreier JL, Lidington D, Bolz SS, Friedman TB, Hla T, Proia RL
JournalJ Biol Chem
PubMed ID17284444
The S1P(2) receptor is a member of a family of G protein-coupled receptors that bind the extracellular sphingolipid metabolite sphingosine 1-phosphate with high affinity. The receptor is widely expressed and linked to multiple G protein signaling pathways, but its physiological function has remained elusive. Here we have demonstrated that S1P(2) ... More
Saccharomyces cerevisiae Hog1 protein phosphorylation upon exposure to bacterial endotoxin.
AuthorsMarques JM, Rodrigues RJ, de Magalhães-Sant'ana AC, Gonçalves T
JournalJ Biol Chem
PubMed ID16790423
The yeast Hog1 protein is both functionally and structurally similar to the mammalian p38, belonging to the same family of mitogen-activated protein (MAP) kinases and responding to extracellular changes in osmolarity. Since p38 mediates lipopolysaccharide (LPS) effects in mammalian cells, we now tested the responsiveness of Hog1 upon exposure of ... More
Expression and localization of Na(+)-HCO(3)(-) cotransporter in bovine corneal endothelium.
AuthorsSun XC, Bonanno JA, Jelamskii S, Xie Q
JournalAm J Physiol Cell Physiol
PubMed ID11029313
Functional studies support the presence of the Na(+)-HCO(3)(-) cotransporter (NBC) in corneal endothelium and possibly corneal epithelium; however, molecular identification and membrane localization have not been reported. To test whether NBC is expressed in bovine cornea, Western blotting was performed, which showed a single band at approximately 130 kDa for ... More
The tumor-sensitive calmodulin-like protein is a specific light chain of human unconventional myosin X.
AuthorsRogers MS, Strehler EE
JournalJ Biol Chem
PubMed ID11278607
Human calmodulin-like protein (CLP) is an epithelial-specific Ca(2+)-binding protein whose expression is strongly down-regulated in cancers. Like calmodulin, CLP is thought to regulate cellular processes via Ca(2+)-dependent interactions with specific target proteins. Using gel overlays, we identified a approximately 210-kDa protein binding specifically and in a Ca(2+)-dependent manner to CLP, ... More
A bipartite Ca2+-regulated nucleoside-diphosphate kinase system within the Chlamydomonas flagellum. The regulatory subunit p72.
AuthorsPatel-King RS, Benashski SE, King SM
JournalJ Biol Chem
PubMed ID12095989
Regulation of flagellar activity in Chlamydomonas involves both Ca(2+) and cAMP-mediated signaling pathways. However, Chlamydomonas and sea urchin sperm flagella also exhibit nucleoside-diphosphate kinase (NDK) activity, suggesting a requirement for GTP within this highly conserved organelle. In sea urchin sperm, the NDK catalytic subunit is an integral component of the ... More
Transglutaminase 5 cross-links loricrin, involucrin, and small proline-rich proteins in vitro.
AuthorsCandi E, Oddi S, Terrinoni A, Paradisi A, Ranalli M, Finazzi-Agró A, Melino G
JournalJ Biol Chem
PubMed ID11443109
Transglutaminases (TGases) are seven enzymes, cross-linking proteins by gamma-glutamil-epsilon-lysine bonds, four of which are expressed in the skin. A new member of the TGase family, TGase 5, has been identified recently, and in the present study we evaluated its role in keratinocyte differentiation in vitro. In addition to the previously ... More