The vomeronasal receptor V2R2 does not require escort molecules for expression in heterologous systems.
AuthorsSilvotti L, Giannini G, Tirindelli R,
JournalChem Senses
PubMed ID15647459
'In rodents, many behavioural responses are triggered by pheromones. These molecules are believed to bind and activate two families of G-protein coupled receptors, namely V1Rs and V2Rs, which are specifically expressed in the chemosensory neurons of the vomeronasal organ. V2Rs are homologous with Group 3 of G-protein-coupled receptors, which includes ... More
Identification of Novel SH3 Domain Ligands for the Src Family Kinase Hck. WISKOTT-ALDRICH SYNDROME PROTEIN (WASP), WASP-INTERACTING PROTEIN (WIP), AND ELMO1.
Authors Scott Margaret Porter; Zappacosta Francesca; Kim Eun Young; Annan Roland S; Miller W Todd;
JournalJ Biol Chem
PubMed ID12029088
'The importance of the SH3 domain of Hck in kinase regulation, substrate phosphorylation, and ligand binding has been established. However, few in vivo ligands are known for the SH3 domain of Hck. In this study, we used mass spectrometry to identify approximately 25 potential binding partners for the SH3 domain ... More
Allocation of helper T-cell epitope immunodominance according to three-dimensional structure in the human immunodeficiency virus type I envelope glycoprotein gp120.
Authors Dai G; Steede N K; Landry S J;
JournalJ Biol Chem
PubMed ID11551929
'The specificity and intensity of CD4(+) helper T-cell responses determine the effectiveness of immune effector functions. Promiscuously immunodominant helper T-cell epitopes in the human immunodeficiency virus (HIV) envelope glycoprotein gp120 could be important in the development of broadly protective immunity, but the underlying mechanisms of immunodominance and promiscuity remain poorly ... More
Expression, purification, and characterization of an activated cytokine-suppressive anti-inflammatory drug-binding protein 2 (CSBP2) kinase from baculovirus-infected insect cells.
Authors Cai X Y; Shanahan M; Miller K; Gommoll C; Lundell D; Zavodny P; Dalie B;
JournalProtein Expr Purif
PubMed ID9226723
'An activated form of the human cytokine-suppressive anti-inflammatory drug-binding protein 2 (CSBP2) kinase was expressed in Spodoptera frugiperda (SF9) cells from a baculovirus vector. To maximize expression and to facilitate purification of the recombinant protein, CSBP2 kinase was expressed as a carboxy-terminal fusion protein to glutathione S-transferase (GST). Under optimal ... More
Kinesin Superfamily Motor Protein KIF17 and mLin-10 in NMDA Receptor-Containing Vesicle Transport
'Experiments with vesicles containing N-methyl-D-aspartate (NMDA)receptor 2B (NR2B subunit) show that they are transported alongmicrotubules by KIF17, a neuron-specific molecular motor in neuronaldendrites. Selective transport is accomplished by direct interaction of theKIF17 tail with a PDZ domain of mLin-10 (Mint1/X11), which is aconstituent of a large protein complex including mLin-2 ... More
Variola virus immune evasion design: expression of a highly efficient inhibitor of human complement.
Authors Rosengard Ariella M; Liu Yu; Nie Zhiping; Jimenez Robert;
JournalProc Natl Acad Sci U S A
PubMed ID12077314
'Variola virus, the most virulent member of the genus Orthopoxvirus, specifically infects humans and has no other animal reservoir. Variola causes the contagious disease smallpox, which has a 30-40% mortality rate. Conversely, the prototype orthopoxvirus, vaccinia, causes no disease in immunocompetent humans and was used in the global eradication of ... More
Purification, cDNA cloning, and expression of UDP-Gal: glucosylceramide beta-1,4-galactosyltransferase from rat brain.
'Lactosylceramide synthase is an enzyme that catalyzes the transfer of galactose from UDP-Gal to glucosylceramide, and thus participates in the biosynthesis of most glycosphingolipids in mammals. We purified this enzyme over 61,000-fold to near homogeneity with a 29. 7% yield from rat brain membrane fractions. The isolation procedure included solubilization ... More
Evidence for a Functional Interaction between Cingulin and ZO-1 in Cultured Cells.
Authors D'Atri Fabio; Nadalutti Fabio; Citi Sandra;
JournalJ Biol Chem
PubMed ID12023291
'Cingulin, a protein component of the submembrane plaque of tight junctions (TJ), contains globular and coiled-coil domains and interacts in vitro with several TJ and cytoskeletal proteins, including the PDZ protein ZO-1. Overexpression of Xenopus cingulin in transfected Xenopus A6 cells resulted in the disruption of endogenous ZO-1 localization, suggesting ... More
The 5-hydroxytryptamine(4a) receptor is palmitoylated at two different sites, and acylation is critically involved in regulation of receptor constitutive activity.
'We have reported recently that the mouse 5-hydroxytryptamine(4a) (5-HT(4(a))) receptor undergoes dynamic palmitoylation (Ponimaskin, E. G., Schmidt, M. F., Heine, M., Bickmeyer, U., and Richter, D. W. (2001) Biochem. J. 353, 627-663). In the present study, conserved cysteine residues 328/329 in the carboxyl terminus of the 5-HT(4(a)) receptor were identified ... More
The major conformational IgE-binding epitopes of hevein (Hev b6.02) are identified by a novel chimera-based allergen epitope mapping strategy.
Authors Karisola Piia; Alenius Harri; Mikkola Jari; Kalkkinen Nisse; Helin Jari; Pentikäinen Olli T; Repo Susanna; Reunala Timo; Turjanmaa Kristiina; Johnson Mark S; Palosuo Timo; Kulomaa Markku S;
JournalJ Biol Chem
PubMed ID11909866
'A novel approach to localize and reconstruct conformational IgE-binding epitope regions of hevein (Hev b6.02), a major natural rubber latex allergen, is described. An antimicrobial protein (AMP) from the amaranth Amaranthus caudatus was used as an immunologically non-IgE-binding adaptor molecule to which terminal or central parts of hevein were fused. ... More
Interaction of Fibroblast Growth Factor Receptor 3 and the Adapter Protein SH2-B. A ROLE IN STAT5 ACTIVATION.
Authors Kong Monica; Wang Ching S; Donoghue Daniel J;
JournalJ Biol Chem
PubMed ID11827956
'Fibroblast growth factor receptor 3 (FGFR3) influences a diverse array of biological processes, including cell growth, differentiation, and migration. Activating mutations in FGFR3 are associated with multiple myeloma, cervical carcinoma, and bladder cancer. To identify proteins that interact with FGFR3 and which may mediate FGFR3-dependent signaling, a yeast two-hybrid screen ... More
Reuptake of extracellular amelogenin by dental epithelial cells results in increased levels of amelogenin mRNA through enhanced mRNA stabilization.
'Amelogenin is an extracellular matrix protein secreted by ameloblasts and is a major component of enamel matrix. Recently, in addition to their role in enamel formation, the biological activity of enamel proteins in the process of cell differentiation has recently become widely appreciated. In this study, we examined the biological ... More
Protein kinase C (PKC)eta-mediated PKC mu activation modulates ERK and JNK signal pathways.
'Protein kinase C (PKC), a family of lipid-activated serine kinases, is involved in multiple functions in the regulation of growth control. The PKC-related isoform PKC mu/PKD has been implicated in mitogenic signal cascades because of the activation of p42/p44 MAPK leading to Elk1-mediated gene transcription, and PKC mu/PKD has been ... More
SCAP ligands are potent new lipid-lowering drugs.
Authors Grand-Perret T; Bouillot A; Perrot A; Commans S; Walker M; Issandou M;
JournalNat Med
PubMed ID11726962
'Upregulation of low-density lipoprotein receptor (LDLr) is a key mechanism to control elevated plasma LDL-cholesterol levels. Here we identify a new class of compounds that directly binds to the sterol regulatory element-binding protein (SREBP) cleavage-activating protein (SCAP). We show that a 14C-labeled, photo-activatable analog specifically labeled both SCAP and a ... More
High incidence of epithelial cancers in mice deficient for DNA polymerase delta proofreading.
Authors Goldsby Robert E; Hays Laura E; Chen Xin; Olmsted Elise A; Slayton William B; Spangrude Gerry J; Preston Bradley D;
JournalProc Natl Acad Sci U S A
PubMed ID12429860
'Mutations are a hallmark of cancer. Normal cells minimize spontaneous mutations through the combined actions of polymerase base selectivity, 3'' --> 5'' exonucleolytic proofreading, mismatch correction, and DNA damage repair. To determine the consequences of defective proofreading in mammals, we created mice with a point mutation (D400A) in the proofreading ... More
15-Lipoxygenase metabolism of 2-arachidonylglycerol. Generation of a peroxisome proliferator-activated receptor alpha agonist.
Authors Kozak Kevin R; Gupta Rajnish A; Moody John S; Ji Chuan; Boeglin William E; DuBois Raymond N; Brash Alan R; Marnett Lawrence J;
JournalJ Biol Chem
PubMed ID11956198
'The recent demonstrations that cyclooxygenase-2 and leukocyte-type 12-lipoxygenase (LOX) efficiently oxygenate 2-arachidonylglycerol (2-AG) prompted an investigation into related oxygenases capable of metabolizing this endogenous cannabinoid receptor ligand. We evaluated the ability of six LOXs to catalyze the hydroperoxidation of 2-AG. Soybean 15-LOX, rabbit reticulocyte 15-LOX, human 15-LOX-1, and human 15-LOX-2 ... More
Characterization of tumor-associated Chk2 mutations.
AuthorsWu X, Webster SR, Chen J,
JournalJ Biol Chem
PubMed ID11053450
'The integrity of the DNA damage response pathway is essential for prevention of neoplastic transformation. Several proteins involved in this pathway including p53, BRCA1, and ATM are frequently mutated in human cancer. Checkpoint kinase 2 (Chk2) is a DNA damage-activated protein kinase that lies downstream of ATM in this pathway. ... More
Preferential methylation of unmethylated DNA by Mammalian de novo DNA methyltransferase dnmt3a.
Authors Yokochi Tomoki; Robertson Keith D;
JournalJ Biol Chem
PubMed ID11821381
'DNA methylation is an epigenetic modification of DNA. There are currently three catalytically active mammalian DNA methyltransferases, DNMT1, -3a, and -3b. DNMT1 has been shown to have a preference for hemimethylated DNA and has therefore been termed the maintenance methyltransferase. Although previous studies on DNMT3a and -3b revealed that they ... More
Evolution of moth sex pheromones via ancestral genes.
Authors Roelofs Wendell L; Liu Weitian; Hao Guixia; Jiao Hongmei; Rooney Alejandro P; Linn Charles E Jr;
JournalProc Natl Acad Sci U S A
PubMed ID12374851
'Mate finding in most moth species involves long-distance signaling via female-emitted sex pheromones. There is a great diversity of pheromone structures used throughout the Lepidoptera, even among closely related species. The conundrum is how signal divergence has occurred. With strong normalizing selection pressure on blend composition and response preferences, it ... More
5-Hydroxytryptamine 4(a) receptor is coupled to the Galpha subunit of heterotrimeric G13 protein.
'Serotonin (5-hydroxytryptamine (5-HT)) is an important neurotransmitter that regulates multiple events in the central nervous system. Many of the 5-HT functions are mediated via G protein-coupled receptors that are coupled to multiple heterotrimeric G proteins, including G(s), G(i), and G(q) subfamilies (Martin, G. R., Eglen, R. M., Hamblin, M. W., ... More
Species specificity of simian virus 40 DNA replication in vitro requires multiple functions of human DNA polymerase alpha.
Authors Smith Richard W P; Steffen Claudia; Grosse Frank; Nasheuer Heinz-Peter;
JournalJ Biol Chem
PubMed ID11927598
'Human cell extracts support the replication of SV40 DNA, whereas mouse cell extracts do not. Species specificity is determined at the level of initiation of DNA replication, and it was previously found that this requires the large subunit, p180, of DNA polymerase alpha-primase to be of human origin. Furthermore, a ... More
DNA chain length dependence of formation and dynamics of hMutSalpha.hMutLalpha.heteroduplex complexes.
Authors Blackwell L J; Wang S; Modrich P;
JournalJ Biol Chem
PubMed ID11441019
'Formation of a ternary complex between human MutSalpha, MutLalpha, and heteroduplex DNA has been demonstrated by surface plasmon resonance spectroscopy and electrophoretic gel shift methods. Formation of the hMutLalpha.hMutSalpha.heteroduplex complex requires a mismatch and ATP hydrolysis, and depends on DNA chain length. Ternary complex formation was supported by a 200-base ... More
Identification of FEZ1 as a protein that interacts with JC virus agnoprotein and microtubules: role of agnoprotein-induced dissociation of FEZ1 from microtubules in viral propagation.
AuthorsSuzuki T, Okada Y, Semba S, Orba Y, Yamanouchi S, Endo S, Tanaka S, Fujita T, Kuroda S, Nagashima K, Sawa H,
JournalJ Biol Chem
PubMed ID15843383
'The human polyomavirus JC virus (JCV) is the causative agent of a fatal demyelinating disease, progressive multifocal leukoencephalopathy, and encodes six major proteins, including agnoprotein. Agnoprotein colocalizes with microtubules in JCV-infected cells, but its function is not fully understood. We have now identified fasciculation and elongation protein zeta 1 (FEZ1) ... More
Enhanced production of green fluorescent fusion proteins in a baculovirus expression system by addition of secretion signal.
AuthorsKatagiri Y, Ingham KC.
JournalBiotechniques
PubMed ID12139250
Structure of the reovirus membrane-penetration protein, Mu1, in a complex with is protector protein, Sigma3.
Authors Liemann Susanne; Chandran Kartik; Baker Timothy S; Nibert Max L; Harrison Stephen C;
JournalCell
PubMed ID11832217
Cell entry by nonenveloped animal viruses requires membrane penetration without membrane fusion. The reovirus penetration agent is the outer-capsid protein, Mu1. The structure of Mu1, complexed with its ... More
Counting low-copy number proteins in a single cell.
We have designed a microfluidic device in which we can manipulate, lyse, label, separate, and quantify the protein contents of a single cell using single-molecule fluorescence counting. Generic labeling of proteins is achieved through fluorescent-antibody binding. The use of cylindrical optics enables high-efficiency (approximately 60%) counting of molecules in micrometer-sized ... More
The nephroblastoma overexpressed gene (NOV/ccn3) protein associates with Notch1 extracellular domain and inhibits myoblast differentiation via Notch signaling pathway.
AuthorsSakamoto K, Yamaguchi S, Ando R, Miyawaki A, Kabasawa Y, Takagi M, Li CL, Perbal B, Katsube K.
JournalJ Biol Chem
PubMed ID12050162
We demonstrate a novel interaction of the nephroblastoma overexpressed gene (NOV), a member of the CCN gene family, with the Notch signaling pathway. NOV associates with the epidermal growth factor-like repeats of Notch1 by the CT (C-terminal cysteine knot) domain. The promoters of HES1 and HES5, which are the downstream ... More
A role for presenilin-1 in nuclear accumulation of Ire1 fragments and induction of the mammalian unfolded protein response.
AuthorsNiwa M, Sidrauski C, Kaufman RJ, Walter P,
JournalCell
PubMed ID10619423
The unfolded protein response (UPR) mediates signaling from the endoplasmic reticulum to the nucleus. In yeast, a key regulatory step in the UPR is the spliceosome-independent splicing of HAC1 mRNA encoding a UPR-specific transcription factor, which is initiated by the transmembrane kinase/endoribonuclease Ire1. We show that yeast HAC1 mRNA is ... More
Involvement of a bifunctional fatty-acyl desaturase in the biosynthesis of the silkmoth, Bombyx mori, sex pheromone.
The straight-chain C(10) to C(18) unsaturated aliphatic compounds containing an oxygenated functional group (aldehyde, alcohol, or acetate ester) derived from saturated C(16) or C(18) fatty acids are a major class of sex pheromone components produced by female moths. In the biosynthesis of these pheromone components, various combinations of limited chain-shortening ... More
Distinct sites on G protein beta gamma subunits regulate different effector functions.
AuthorsMirshahi T, Mittal V, Zhang H, Linder ME, Logothetis DE,
JournalJ Biol Chem
PubMed ID12124391
G proteins interact with effectors at multiple sites and regulate their activity. The functional significance of multiple contact points is not well understood. We previously identified three residues on distinct surfaces of Gbetagamma that are crucial for G protein-coupled inward rectifier K(+) (GIRK) channel activation. Here we show that mutations ... More
PIAS1 and PIASxalpha function as SUMO-E3 ligases toward androgen receptor and repress androgen receptor-dependent transcription.
AuthorsNishida T, Yasuda H,
JournalJ Biol Chem
PubMed ID12177000
The androgen receptor (AR) has been shown to be modified by SUMO-1, a ubiquitin-like protein. Recently we showed that PIAS family proteins function as SUMO-E3 ligases. Here we provide evidence that PIAS1 and PIASxalpha act as specific SUMO-E3 ligases for the AR. PIAS1 and PIASxalpha but not PIAS3 or PIASxbeta ... More
High-efficiency transient transduction of human embryonic stem cell-derived neurons with baculoviral vectors.
AuthorsZeng J, Du J, Lin J, Bak XY, Wu C, Wang S,
JournalMol Ther
PubMed ID19532141
Transient genetic manipulation of human neurons without chromosomal integration of the transgene would be valuable but has been challenging due to the quiescent nature of these postmitotic cells. In this study, we developed a set of baculoviral vectors for transient transduction in nondividing neurons derived from human embryonic stem cells ... More
Characterization of a heparan sulfate 3-O-sulfotransferase-5, an enzyme synthesizing a tetrasulfated disaccharide.
AuthorsMochizuki H, Yoshida K, Gotoh M, Sugioka S, Kikuchi N, Kwon YD, Tawada A, Maeyama K, Inaba N, Hiruma T, Kimata K, Narimatsu H,
JournalJ Biol Chem
PubMed ID12740361
Heparan sulfate d-glucosaminyl 3-O-sulfotransferases (3-OSTs) catalyze the transfer of sulfate from 3'-phosphoadenosine 5'-phosphosulfate (PAPS) to position 3 of the glucosamine residue of heparan sulfate and heparin. A sixth member of the human 3-OST family, named 3-OST-5, was recently reported (Xia, G., Chen, J., Tiwari, V., Ju, W., Li, J.-P., Malmstrom, ... More
Structure and conformational changes in the C-terminal domain of the beta2-adrenoceptor: insights from fluorescence resonance energy transfer studies.
The C terminus of the beta(2)-adrenoceptor (AR) interacts with G protein-coupled receptor kinases and arrestins in an agonist-dependent manner, suggesting that conformational changes induced by ligands in the transmembrane domains are transmitted to the C terminus. We used fluorescence resonance energy transfer (FRET) to examine ligand-induced structural changes in the ... More
All Six Modules of the Gelatin-binding Domain of Fibronectin Are Required for Full Affinity.
AuthorsKatagiri Y, Brew SA, Ingham KC,
JournalJ Biol Chem
PubMed ID12538576
The gelatin-binding sites of fibronectin are confined to a 42-kDa region having four type I and two type II modules in the following order: I(6)-II(1)-II(2)-I(7)-I(8)-I(9). To determine the relative importance of each module for recognition of gelatin, recombinant green fluorescent fusion proteins were prepared in which individual modules or groups ... More
The UL12.5 gene product of herpes simplex virus type 1 exhibits nuclease and strand exchange activities but does not localize to the nucleus.
AuthorsReuven NB, Antoku S, Weller SK,
JournalJ Virol
PubMed ID15078942
The herpes simplex virus type 1 (HSV-1) alkaline nuclease, encoded by the UL12 gene, plays an important role in HSV-1 replication, as a null mutant of UL12 displays a severe growth defect. Although the precise in vivo role of UL12 has not yet been determined, several in vitro activities have ... More
Murine and human autotaxin alpha, beta, and gamma isoforms: gene organization, tissue distribution, and biochemical characterization.
AuthorsGiganti A, Rodriguez M, Fould B, Moulharat N, Cogé F, Chomarat P, Galizzi JP, Valet P, Saulnier-Blache JS, Boutin JA, Ferry G,
JournalJ Biol Chem
PubMed ID18175805
Autotaxin is a type II ectonucleotide pyrophosphate phosphodiesterase enzyme. It has been recently discovered that it also has a lysophospholipase D activity. This enzyme probably provides most of the extracellular lysophosphatidic acid from lysophosphatidylcholine. The cloning and tissue distribution of the three isoforms (imaginatively called alpha, beta, and gamma) from ... More
Mechanism of Ca2+ disruption in Alzheimer's disease by presenilin regulation of InsP(3) receptor channel gating.
AuthorsCheung KH, Shineman D, Müller M, Cárdenas C, Mei L, Yang J, Tomita T, Iwatsubo T, Lee VM, Foskett JK,
JournalNeuron
PubMed ID18579078
Mutations in presenilins (PS) are the major cause of familial Alzheimer's disease (FAD) and have been associated with calcium (Ca2+) signaling abnormalities. Here, we demonstrate that FAD mutant PS1 (M146L)and PS2 (N141I) interact with the inositol 1,4,5-trisphosphate receptor (InsP3R) Ca2+ release channel and exert profound stimulatory effects on its gating ... More
The novel human DNA helicase hFBH1 is an F-box protein.
Authors Kim Jaehoon; Kim Jeong-Hoon; Lee Sung-Hak; Kim Do-Hyung; Kang Ho-Young; Bae Sung-Ho; Pan Zhen-Qiang; Seo Yeon-Soo;
JournalJ Biol Chem
PubMed ID11956208
We have identified a novel DNA helicase in humans that belongs to members of the superfamily I helicase and found that it contains a well conserved F-box motif at its N terminus. We have named the enzyme hFBH1 (human F-box DNA helicase 1). Recombinant hFBH1, containing glutathione S-transferase at the ... More
Human MutY homolog, a DNA glycosylase involved in base excision repair, physically and functionally interacts with mismatch repair proteins human MutS homolog 2/human MutS homolog 6.
Authors Gu Yesong; Parker Antony; Wilson Teresa M; Bai Haibo; Chang Dau-Yin; Lu A-Lien;
JournalJ Biol Chem
PubMed ID11801590
Adenines mismatched with guanines or 7,8-dihydro-8-oxo-deoxyguanines that arise through DNA replication errors can be repaired by either base excision repair or mismatch repair. The human MutY homolog (hMYH), a DNA glycosylase, removes adenines from these mismatches. Human MutS homologs, hMSH2/hMSH6 (hMutSalpha), bind to the mismatches and initiate the repair on ... More
Elimination of P1 arginine 393 interaction with underlying glutamic acid 255 partially activates antithrombin III for thrombin inhibition but not factor Xa inhibition.
Authors Jairajpuri Mohamad Aman; Lu Aiqin; Bock Susan C;
JournalJ Biol Chem
PubMed ID11971909
The mechanism for heparin activation of antithrombin III has been postulated to involve disruption of interactions between its reactive loop P1 residue and Glu(255) on the underlying protein surface. To test this hypothesis, the potential P1-constraining Arg(393)-Glu(255) hydrogen bond and ionic interactions were eliminated by converting Glu(255) to alanine. E255A ... More
CR16 forms a complex with N-WASP in brain and is a novel member of a conserved proline-rich actin-binding protein family.
Authors Ho H Y; Rohatgi R; Ma L; Kirschner M W;
JournalProc Natl Acad Sci U S A
PubMed ID11553796
The Neuronal Wiskott-Aldrich syndrome protein (N-WASP) has emerged as a central regulator of the actin cytoskeleton with abilities to integrate multiple upstream signal inputs and transmit them to the Arp2/3 complex. Here, we demonstrate that native N-WASP is present in a tight complex with a proline-rich protein, CR16, which shares ... More
Dissociation time from DNA determines transcriptional function in a STAT1 linker mutant.
Authors Yang Edward; Henriksen Melissa A; Schaefer Olaf; Zakharova Natalia; Darnell James E Jr;
JournalJ Biol Chem
PubMed ID11834743
The STAT1 transcription factor is organized into several highly conserved domains, each of which has been assigned a function with the exception of the linker domain. We previously characterized a mutant in the linker domain of STAT1 that gave normal DNA binding using a standard probe in an electrophoretic mobility ... More
Functional analysis of mutant and wild-type Drosophila origin recognition complex.
Authors Chesnokov I; Remus D; Botchan M;
JournalProc Natl Acad Sci U S A
PubMed ID11593009
The origin recognition complex (ORC) is the DNA replication initiator protein in eukaryotes. We have reconstituted a functional recombinant Drosophila ORC and compared activities of the wild-type and several mutant ORC variants. Drosophila ORC is an ATPase, and our studies show that the ORC1 subunit is essential for ATP hydrolysis ... More
Expression, purification, and characterization of gp160e, the soluble, trimeric ectodomain of the simian immunodeficiency virus envelope glycoprotein, gp160.
Authors Chen B; Zhou G; Kim M; Chishti Y; Hussey R E; Ely B; Skehel J J; Reinherz E L; Harrison S C; Wiley D C;
JournalJ Biol Chem
PubMed ID10944528
The envelope glycoprotein, gp160, of simian immunodeficiency virus (SIV) shares approximately 25% sequence identity with gp160 from the human immunodeficiency virus, type I, indicating a close structural similarity. As a result of binding to cell surface CD4 and co-receptor (e.g. CCR5 and CXCR4), both SIV and human immunodeficiency virus gp160 ... More
Human exonuclease I is required for 5' and 3' mismatch repair.
Authors Genschel Jochen; Bazemore Laura R; Modrich Paul;
JournalJ Biol Chem
PubMed ID11809771
We have partially purified a human activity that restores mismatch-dependent, bi-directional excision to a human nuclear extract fraction depleted for one or more mismatch repair excision activities. Human EXOI co-purifies with the excision activity, and the purified activity can be replaced by near homogeneous recombinant hEXOI. Despite the reported 5' ... More
The delta subunit of retinal rod cGMP phosphodiesterase regulates the membrane association of Ras and Rap GTPases.
Authors Nancy Vanessa; Callebaut Isabelle; El Marjou Ahmed; de Gunzburg Jean;
JournalJ Biol Chem
PubMed ID11786539
Post-translational modifications of GTPases from the Ras superfamily enable them to associate with membrane compartments where they exert their biological activities. However, no protein acting like Rho and Rab dissociation inhibitor (GDI) that regulate the membrane association of Rho and Rab GTPases has been described for Ras and closely related ... More
Substrate and inhibitor profile of BACE (beta-secretase) and comparison with other mammalian aspartic proteases.
The full-length and ectodomain forms of beta-site APP cleavage enzyme (BACE) have been cloned, expressed in Sf9 cells, and purified to homogeneity. This aspartic protease cleaves the amyloid precursor protein at the beta-secretase site, a critical step in the Alzheimer's disease pathogenesis. Comparison of BACE to other aspartic proteases such ... More
Autoinhibition of the ligand-binding site of GGA1/3 VHS domains by an internal acidic cluster-dileucine motif.
Authors Doray Balraj; Bruns Kerry; Ghosh Pradipta; Kornfeld Stuart A;
JournalProc Natl Acad Sci U S A
PubMed ID12060753
The GGAs (Golgi-localizing, gamma-adaptin ear homology domain, ARF-binding proteins) are a family of proteins implicated in protein trafficking from the Golgi to endosomes/lysosomes. These proteins have modular structures with an N-terminal VHS (VPS-27, Hrs, and STAM) domain followed by a GAT (GGA and TOM1) domain, a connecting hinge segment, and ... More
Biochemical properties of purified recombinant human beta-carotene 15,15'-monooxygenase.
Authors Lindqvist Annika; Andersson Stefan;
JournalJ Biol Chem
PubMed ID11960992
Beta-carotene 15,15'-monooxygenase (BCO), formerly known as beta-carotene 15,15'-dioxygenase, catalyzes the first step in the synthesis of vitamin A from dietary carotenoids. We have biochemically and enzymologically characterized the purified recombinant human BCO enzyme. A highly active BCO enzyme was expressed and purified to homogeneity from baculovirus-infected Spodoptera frugiperda 9 insect ... More
The Influence of the Cdc27 Subunit on the Properties of the Schizosaccharomyces pombe DNA Polymerase delta.
Authors Bermudez Vladimir P; MacNeill Stuart A; Tappin Inger; Hurwitz Jerard;
JournalJ Biol Chem
PubMed ID12124382
Schizosaccharomyces pombe DNA polymerase (pol) delta contains four subunits, pol 3, Cdc1, Cdc27, and Cdm1. In this report, we examined the role of Cdc27 on the structure and activity of pol delta. We show that the four-subunit complex is monomeric in structure, in contrast to the previous report that it ... More
Comparison of filamin A-induced cross-linking and Arp2/3 complex-mediated branching on the mechanics of actin filaments.
Authors Nakamura Fumihiko; Osborn Eric; Janmey Paul A; Stossel Thomas P;
JournalJ Biol Chem
PubMed ID11786548
We compared the effects of human filamin A (FLNa) and the activated human Arp2/3 complex on mechanical properties of actin filaments. As little as 1 FLNa to 800 polymerizing actin monomers induces a sharp concentration-dependent increase in the apparent viscosity of 24 microm actin, a parameter classically defined as a ... More
JAM2 Interacts with alpha 4beta 1. FACILITATION BY JAM3.
Authors Cunningham Sonia A; Rodriguez Jose M; Arrate M Pia; Tran Tuan M; Brock Tommy A;
JournalJ Biol Chem
PubMed ID12070135
We have previously reported that junctional adhesion molecule 2 (JAM2) adheres to T cells through heterotypic interactions with JAM3. An examination of the cation dependence of JAM2 adhesion to HSB cells revealed a Mn(2+)-enhanced binding component indicative of integrin involvement. Using neutralizing integrin antibodies, we have defined an interaction between ... More
Comparison of Cak1p-like cyclin-dependent kinase-activating kinases.
Authors Tsakraklides Vasiliki; Solomon Mark J;
JournalJ Biol Chem
PubMed ID12084729
Cyclin-dependent kinases (cdks) coordinate progression through the eukaryotic cell cycle and require phosphorylation by a cdk-activating kinase (CAK) for full activity. In most eukaryotes Cdk7 is the catalytic subunit of a heterotrimeric CAK (Cdk7-cyclin H-Mat1) that is also involved in transcription as part of the transcription factor IIH complex. The ... More
Tetrahymena proteins p80 and p95 are not core telomerase components.
Authors Mason D X; Autexier C; Greider C W;
JournalProc Natl Acad Sci U S A
PubMed ID11592988
Telomeres provide stability to eukaryotic chromosomes and consist of tandem DNA repeat sequences. Telomeric repeats are synthesized and maintained by a specialized reverse transcriptase, termed telomerase. Tetrahymena thermophila telomerase contains two essential components: Tetrahymena telomerase reverse transcriptase (tTERT), the catalytic protein component, and telomerase RNA that provides the template for ... More
Ubc9 is a novel modulator of the induction properties of glucocorticoid receptors.
Authors Kaul Sunil; Blackford John A Jr; Cho Sehyung; Simons S Stoney Jr;
JournalJ Biol Chem
PubMed ID11812797
The EC(50) of agonists and the partial agonist activity of antagonists are crucial parameters for steroid hormone control of gene expression and endocrine therapies. These parameters have been shown to be modulated by a naturally occurring cis-acting element, called the glucocorticoid modulatory element (GME) that binds two proteins, GMEB-1 and ... More
The (I/Y)XGG motif of adenovirus DNA polymerase affects template DNA binding and the transition from initiation to elongation.
Authors Brenkman A B; Heideman M R; Truniger V; Salas M; van der Vliet P C;
JournalJ Biol Chem
PubMed ID11390396
Adenovirus DNA polymerase (Ad pol) is a eukaryotic-type DNA polymerase involved in the catalysis of protein-primed initiation as well as DNA polymerization. The functional significance of the (I/Y)XGG motif, highly conserved among eukaryotic-type DNA polymerases, was analyzed in Ad pol by site-directed mutagenesis of four conserved amino acids. All mutant ... More
p53 recruitment of CREB binding protein mediated through phosphorylated CREB: a novel pathway of tumor suppressor regulation.
Authors Giebler H A; Lemasson I; Nyborg J K;
JournalMol Cell Biol
PubMed ID10848610
CREB binding protein (CBP) is a 270-kDa nuclear protein required for activated transcription of a large number of cellular genes. Although CBP was originally discovered through its interaction with phosphorylated CREB (pCREB), it is utilized by a multitude of cellular transcription factors and viral oncoproteins. Both CREB and the tumor ... More
Molecular cloning and characterization of a novel UDP-GlcNAc:GalNAc-peptide beta1,3-N-acetylglucosaminyltransferase (beta 3Gn-T6), an enzyme synthesizing the core 3 structure of O-glycans.
The core 3 structure of the O-glycan, GlcNAcbeta1-3GalNAcalpha1-serine/threonine, an important precursor in the biosynthesis of mucin-type glycoproteins, is synthesized by UDP-N-acetylglucosamine:GalNAc-peptide beta1,3-N- acetylglucosaminyltransferase (beta3Gn-T; core 3 synthase). The core 3 structure is restricted in its occurrence to mucins from specific tissues such as the stomach, small intestine, and colon. A ... More
Hedgehog-stimulated Phosphorylation of the Kinesin-related Protein Costal2 Is Mediated by the Serine/Threonine Kinase Fused.
Authors Nybakken Kent E; Turck Christoph W; Robbins David J; Bishop J Michael;
JournalJ Biol Chem
PubMed ID11934882
The Hedgehog (Hh) signaling molecule is required for the development of numerous tissues in Drosophila. Within the cell, Hh signal transduction utilizes a large protein complex consisting of the Fused (Fu), Costal2 (Cos2), and Cubitis interruptus (Ci) proteins, but the functional interactions between these proteins are still largely uncharacterized. Using ... More
Chemical quenched flow kinetic studies indicate an intraholoenzyme autophosphorylation mechanism for Ca2+/calmodulin-dependent protein kinase II.
Autophosphorylation of alpha-Ca(2+)/calmodulin-dependent protein kinase II (CaM kinase II) at Thr-286 generates Ca(2+)-independent activity that outlasts the initial Ca(2+) stimulus. Previous studies suggested that this autophosphorylation occurs between subunits within each CaM kinase II holoenzyme. However, electron microscopy studies have questioned this mechanism because a large distance separates a kinase ... More
Photolabeling of human and murine multidrug resistance protein 1 with the high affinity inhibitor [125I]LY475776 and azidophenacyl-[35S]glutathione.
Authors Qian Yue-Ming; Grant Caroline E; Westlake Christopher J; Zhang Da-Wei; Lander Peter A; Shepard Robert L; Dantzig Anne H; Cole Susan P C; Deeley Roger G;
JournalJ Biol Chem
PubMed ID12138119
Multidrug resistance protein 1 (MRP1/ABCC1) is an ATP-dependent transporter of structurally diverse organic anion conjugates. The protein also actively transports a number of non-conjugated chemotherapeutic drugs and certain anionic conjugates by a presently poorly understood GSH-dependent mechanism. LY475776is a newly developed (125)I-labeled azido tricyclic isoxazole that binds toMRP1 with high ... More
Purification and Characterization of the Schizosaccharomyces pombe Origin Recognition Complex. INTERACTION WITH ORIGIN DNA AND Cdc18 PROTEIN.
Authors Chuang Ray-Yuan; Chretien Louise; Dai Jianli; Kelly Thomas J;
JournalJ Biol Chem
PubMed ID11850415
The origin recognition complex (ORC) plays a central role in the initiation of DNA replication in eukaryotic cells. It interacts with origins of DNA replication in chromosomal DNA and recruits additional replication proteins to form functional initiation complexes. These processes have not been well characterized at the biochemical level except ... More
Identification and characterization of a novel human microsomal glutathione S-transferase with leukotriene C4 synthase activity and significant sequence identity to 5-lipoxygenase-activating protein and leukotriene C4 synthase.
Authors Jakobsson P J; Mancini J A; Ford-Hutchinson A W;
JournalJ Biol Chem
PubMed ID8703034
5-Lipoxygenase-activating protein (FLAP) and leukotriene C4 (LTC4) synthase, two proteins involved in leukotriene biosynthesis, have been demonstrated to be 31% identical at the amino acid level. We have recently identified and characterized a novel member of the FLAP/LTC4 synthase gene family termed microsomal glutathione S-transferase II (microsomal GST-II). The open ... More
Molecular basis for the substrate specificity of NIMA-related kinase-6 (NEK6). Evidence that NEK6 does not phosphorylate the hydrophobic motif of ribosomal S6 protein kinase and serum- and glucocorticoid-induced protein kinase in vivo.
Authors Lizcano Jose M; Deak Maria; Morrice Nick; Kieloch Agnieszka; Hastie C James; Dong Liying; Schutkowski Mike; Reimer Ulf; Alessi Dario R;
JournalJ Biol Chem
PubMed ID12023960
The AGC family of protein kinases, which includes isoforms of protein kinase B (also known as Akt), ribosomal S6 protein kinase (S6K), and serum- and glucocorticoid-induced protein kinase (SGK) are activated in response to many extracellular signals and play key roles in regulating diverse cellular processes. They are activated by ... More
Identification of another actin-related protein (Arp) 2/3 complex binding site in neural Wiskott-Aldrich syndrome protein (N-WASP) that complements actin polymerization induced by the Arp2/3 complex activating (VCA) domain of N-WASP.
Authors Suetsugu S; Miki H; Takenawa T;
JournalJ Biol Chem
PubMed ID11432863
Neural Wiskott-Aldrich syndrome protein (N-WASP) is an essential regulator of actin cytoskeleton formation via its association with the actin-related protein (Arp) 2/3 complex. It is believed that the C-terminal Arp2/3 complex-activating domain (verprolin homology, cofilin homology, and acidic (VCA) or C-terminal region of WASP family proteins domain) of N-WASP is ... More