Activation of calpain I converts excitotoxic neuron death into a caspase-independent cell death.
AuthorsLankiewicz S, Marc Luetjens C, Truc Bui N, Krohn AJ, Poppe M, Cole GM, Saido TC, Prehn JH
JournalJ Biol Chem
PubMed ID10828077
'Glutamate receptor overactivation contributes to neuron death after stroke, trauma, and epileptic seizures. Exposure of cultured rat hippocampal neurons to the selective glutamate receptor agonist N-methyl-d-aspartate (300 microm, 5 min) or to the apoptosis-inducing protein kinase inhibitor staurosporine (300 nm) induced a delayed neuron death. In both cases, neuron death ... More
Luminal pH regulated calcium release kinetics in sarcoplasmic reticulum vesicles.
AuthorsDonoso P, Beltrán M, Hidalgo C
JournalBiochemistry
PubMed ID8873610
'Calcium binding to triads isolated from rabbit skeletal muscle followed a single hyperbolic function in the pH range 5.5-8.0. Maximal binding was obtained at pH 8.0; decreasing the pH decreased the binding capacity and, at pH < or = 6.0, increased Kd 2-fold. These results indicate that lowering the pH ... More
A recombinant tail-less integrin beta 4 subunit disrupts hemidesmosomes, but does not suppress alpha 6 beta 4-mediated cell adhesion to laminins.
AuthorsSpinardi L, Einheber S, Cullen T, Milner TA, Giancotti FG
JournalJ Cell Biol
PubMed ID7721947
'To examine the function of the alpha 6 beta 4 integrin we have determined its ligand-binding ability and overexpressed two potentially dominant negative mutant beta 4 subunits, lacking either the cytoplasmic or extracellular domain, in bladder epithelial 804G cells. The results of cell adhesion and radioligand-binding assays showed that alpha ... More
Crystallization and properties of cathepsin B from rat liver.
AuthorsTowatari T, Kawabata Y, Katunuma N
JournalEur J Biochem
PubMed ID42540
'Cathepsin B from rat liver was purified to apparent homogeneity by cell-fractionation, freezing and thawing, acetone treatment, gel filtration, DEAE-Sephadex and CM-Sephadex column chromatography, and was crystallized. The purified enzyme formed spindle-shaped crystals and its homogeneity was proved by disc gel electrophoresis in the presence of sodium dodecyl sulfate and ... More
Purification, characterization, and localization of follipsin, a novel serine proteinase from the fluid of porcine ovarian follicles.
AuthorsHamabata T, Okimura H, Yokoyama N, Takahashi T, Takahashi K
JournalJ Biol Chem
PubMed ID8027045
'Follipsin, an enzyme that accumulates in the follicular fluid of porcine ovaries during follicular maturation, was purified to apparent homogeneity. The purified enzyme consists of two different polypeptide chains having M(r) = 45,000 and 32,000 each, associated covalently. The enzyme activity was strongly inhibited by diisopropyl fluorophosphate, benzamidine, leupeptin, and ... More
Autocrine motility factor receptor is a marker for a distinct membranous tubular organelle.
AuthorsBenlimame N, Simard D, Nabi IR
JournalJ Cell Biol
PubMed ID7721946
'Autocrine motility factor (AMF) is secreted by tumor cells and is capable of stimulating the motility of the secreting cells. In addition to being expressed on the cell surface, its receptor, AMF-R, is found within a Triton X-100 extractable intracellular tubular compartment. AMF-R tubules can be distinguished by double immunofluorescence ... More
The pH-dependent membrane association of procathepsin L is mediated by a 9-residue sequence within the propeptide.
AuthorsMcIntyre GF, Godbold GD, Erickson AH
JournalJ Biol Chem
PubMed ID8276852
'The lysosomal proprotease procathepsin L binds to mouse fibroblast microsomal membranes at pH 5, but mature active cathepsin L does not (McIntyre, G.F., and Erickson, A. H. (1991) J. Biol. Chem. 266, 15438-15445). This binding is not dependent on N-linked carbohydrate as procathepsin L synthesized in cells treated with tunicamycin ... More
Purification and characterization of two Cl- -activated aminopeptidases hydrolysing basic termini from human skeletal muscle.
'Two aminopeptidases (I and II), hydrolysing basic termini, were purified to homogeneity (as judged by polyacrylamide gel electrophoresis) from human quadriceps muscle by anion-exchange chromatography and preparative electrophoresis. The electrophoretic migration rate of II was approximately 80% of that of I. Both enzymes had the following properties: optimum activity was ... More
Sphingosine-induced apoptosis is dependent on lysosomal proteases.
AuthorsKågedal K, Zhao M, Svensson I, Brunk UT
JournalBiochem J
PubMed ID11583579
'We propose a new mechanism for sphingosine-induced apoptosis, involving relocation of lysosomal hydrolases to the cytosol. Owing to its lysosomotropic properties, sphingosine, which is also a detergent, especially when protonated, accumulates by proton trapping within the acidic vacuolar apparatus, where most of its action as a detergent would be exerted. ... More
Characterization of the insulin-regulated membrane aminopeptidase in 3T3-L1 adipocytes.
AuthorsRoss SA, Scott HM, Morris NJ, Leung WY, Mao F, Lienhard GE, Keller SR
JournalJ Biol Chem
PubMed ID8621739
'A novel membrane aminopeptidase has been identified as a major protein in vesicles from rat adipocytes containing the glucose transporter isotype Glut4. In this study we have characterized this aminopeptidase, referred to as vp165, in 3T3-L1 adipocytes. The subcellular distributions of vp165 and Glut4 were determined by immunoisolation of vesicles ... More
Purification and characterization of DNA ligase III from bovine testes. Homology with DNA ligase II and vaccinia DNA ligase.
'Mammalian cell nuclei contain three biochemically distinct DNA ligases. In the present study we have found high levels of DNA ligase I and DNA ligase III activity in bovine testes and have purified DNA ligase III to near homogeneity. The high level of DNA ligase III suggests a role for ... More
Thioredoxin is required for vacuole inheritance in Saccharomyces cerevisiae.
AuthorsXu Z, Wickner W
JournalJ Cell Biol
PubMed ID8603912
'The vacuole of Saccharomyces cerevisiae projects a stream of tubules a and vesicles (a "segregation structure") into the bud in early S phase. We have described an in vitro reaction, requiring physiological temperature, ATP, and cytosol, in which isolated vacuoles form segregation structures and fuse. This in vitro reaction is ... More
Transferrin-binding protein complex is the receptor for transferrin uptake in Trypanosoma brucei.
AuthorsSteverding D, Stierhof YD, Fuchs H, Tauber R, Overath P
JournalJ Cell Biol
PubMed ID8522581
'In Trypanosoma brucei, the products of two genes, ESAG 6 and ESAG 7, located upstream of the variant surface glycoprotein gene in a polycistronic expression site form a glycosylphosphatidylinositol-anchored transferrin-binding protein (TFBP) complex. It is shown by gel filtration and membrane-binding experiments that the TFBP complex is heterodimeric and binds ... More
Bestatin, an inhibitor of aminopeptidase B, produced by actinomycetes.
AuthorsUmezawa H, Aoyagi T, Suda H, Hamada M, Takeuchi T
JournalJ Antibiot (Tokyo)
PubMed ID931798
Classification of peptidases.
AuthorsBarrett AJ
JournalMethods Enzymol
PubMed ID7845199
Tackling the protease problem in Saccharomyces cerevisiae.
AuthorsJones EW
JournalMethods Enzymol
PubMed ID2005802
Probing the cathepsin D using a BODIPY FL-pepstatin A: applications in fluorescence polarization and microscopy.
AuthorsChen CS, Chen WN, Zhou M, Arttamangkul S, Haugland RP
JournalJ Biochem Biophys Methods
PubMed ID10737220
Redistribution of cathepsin D, a major lysosomal aspartic endopeptidase, has been related to various pathological progressions during tumor formation and oxidation stress. We have synthesized a fluorescent probe for cathepsin D, where the pepstatin A was covalently conjugated with the BODIPY (Boron dipyrromethene difluoride) fluorophore. In vitro, BODIPY FL-pepstatin A ... More
Isolation and characterization of a pepsin C zymogen produced by human breast tissues.
An aspartic proteinase present in cyst fluid from women with gross cystic breast disease was purified by a procedure involving affinity chromatography on pepstatin-agarose and size-exclusion high performance liquid chromatography. The amino-terminal sequence of the purified breast proteinase was identical to that corresponding to gastric pepsinogen C. Additional data on ... More
Calcium-dependent conformation of a mouse macrophage calcium-type lectin. Carbohydrate binding activity is stabilized by an antibody specific for a calcium-dependent epitope.
AuthorsKimura T, Imai Y, Irimura T
JournalJ Biol Chem
PubMed ID7541793
We established monoclonal antibodies (mAbs) against the mouse macrophage galactose/N-acetylgalactosamine-specific lectin (MMGL) that is a 42-kDa calcium-dependent lectin, using a solid phase carbohydrate binding assay as a novel strategy for screening mAbs. The specificity of six mAbs were investigated by antibody binding to native or recombinant forms (rML) of MMGL, ... More
Moesin, ezrin, and p205 are actin-binding proteins associated with neutrophil plasma membranes.
AuthorsPestonjamasp K, Amieva MR, Strassel CP, Nauseef WM, Furthmayr H, Luna EJ
JournalMol Biol Cell
PubMed ID7612961
Actin-binding proteins in bovine neutrophil plasma membranes were identified using blot overlays with 125I-labeled F-actin. Along with surface-biotinylated proteins, membranes were enriched in major actin-binding polypeptides of 78, 81, and 205 kDa. Binding was specific for F-actin because G-actin did not bind. Further, unlabeled F-actin blocked the binding of 125I-labeled ... More
Novel E-cadherin-mediated adhesion in peripheral nerve: Schwann cell architecture is stabilized by autotypic adherens junctions.
AuthorsFannon AM, Sherman DL, Ilyina-Gragerova G, Brophy PJ, Friedrich VL, Colman DR
JournalJ Cell Biol
PubMed ID7698985
Previous studies (Blank, W. F., M. B. Bunge, and R. P. Bunge. 1974. Brain Res. 67:503-518) showed that Schwann cell paranodal membranes were disrupted in calcium free medium suggesting that cadherin mediated mechanisms may operate to maintain the integrity of the paranodal membrane complex. Using antibodies against the fifth extracellular ... More
Cathepsin D as a prognostic indicator for node-negative breast cancer patients using both immunoassays and enzymatic assays.
AuthorsKute TE, Shao ZM, Sugg NK, Long RT, Russell GB, Case LD
JournalCancer Res
PubMed ID1394123
This is a retrospective study on 162 node-negative patients, with both biochemical and clinical factors being measured for determination of prognostic markers. Steroid receptors were measured on all tumors, while tumor size, histological grade, ploidy status, and cell cycle kinetics indicators could not be found or measured on 25 or ... More
The fatty acid bimodal action on superoxide anion production by human adherent monocytes under phorbol 12-myristate 13-acetate or diacylglycerol activation can be explained by the modulation of protein kinase C and p47phox translocation.
AuthorsKadri-Hassani N, Léger CL, Descomps B
JournalJ Biol Chem
PubMed ID7797495
We studied the translocation of protein kinase C (PKC), the endogenous phosphorylation and presence in the membrane fraction of p47phox (the 47-kDa cytosolic component of the phagocyte NADPH oxidase), and the O-.2 production in human adherent monocytes (HAMs). This was performed under phorbol myristate acetate (PMA) or diacylglycerol stimulation after ... More
Crystal structure of human pepsin and its complex with pepstatin.
AuthorsFujinaga M, Chernaia MM, Tarasova NI, Mosimann SC, James MN
JournalProtein Sci
PubMed ID7663352
The three-dimensional crystal structure of human pepsin and that of its complex with pepstatin have been solved by X-ray crystallographic methods. The native pepsin structure has been refined with data collected to 2.2 A resolution to an R-factor of 19.7%. The pepsin:pepstatin structure has been refined with data to 2.0 ... More
Purification and characterization of a collagen-degrading protease from Porphyromonas gingivalis.
AuthorsBedi GS, Williams T
JournalJ Biol Chem
PubMed ID7506259
A trypsin-like protease was purified from spent culture medium of oral pathogen Porphyromonas gingivalis by chromatography on columns of DEAE-Sepharose, gel filtration on Sephadex G-100, and chromatofocusing on PBE-94. Purified enzyme showed a single band on SDS-polyacrylamide gel electrophoresis with an estimated molecular weight of 55,000. Purified protease hydrolyzed type ... More
Alpha-scorpion toxins binding on rat brain and insect sodium channels reveal divergent allosteric modulations by brevetoxin and veratridine.
AuthorsCestèle S, Ben Khalifa RB, Pelhate M, Rochat H, Gordon D
JournalJ Biol Chem
PubMed ID7797499
At least six topologically separated neurotoxin receptor sites have been identified on sodium channels that reveal strong allosteric interactions among them. We have studied the allosteric modulation induced by veratridine, binding to receptor site 2, and brevetoxin PbTx-1, occupying receptor site 5, on the binding of alpha-scorpion toxins at receptor ... More
The VPH1 gene encodes a 95-kDa integral membrane polypeptide required for in vivo assembly and activity of the yeast vacuolar H(+)-ATPase.
AuthorsManolson MF, Proteau D, Preston RA, Stenbit A, Roberts BT, Hoyt MA, Preuss D, Mulholland J, Botstein D, Jones EW
JournalJ Biol Chem
PubMed ID1385813
Yeast vacuolar acidification-defective (vph) mutants were identified using the pH-sensitive fluorescence of 6-carboxyfluorescein diacetate (Preston, R. A., Murphy, R. F., and Jones, E. W. (1989) Proc. Natl. Acad. Sci. U.S.A. 86, 7027-7031). Vacuoles purified from yeast bearing the vph1-1 mutation had no detectable bafilomycin-sensitive ATPase activity or ATP-dependent proton pumping. ... More
Purification, characterization, and conformational analysis of rabbit plasma lipid transfer protein.
AuthorsKo KW, Oikawa K, Ohnishi T, Kay CM, Yokoyama S
JournalBiochemistry
PubMed ID8329397
A procedure for rapid isolation of lipid transfer protein (LTP) from commercially available rabbit plasma is described. Use of protease inhibitors was important for obtaining intact, stable LTP. After lipoproteins were precipitated from the plasma by dextran sulfate, column chromatographies through Butyl-Toyopearl 650M, CM-Toyopearl 650M, and Butyl-Toyopearl 650M were employed. ... More
Inhibition of aspartic proteases by pepstatin and 3-methylstatine derivatives of pepstatin. Evidence for collected-substrate enzyme inhibition.
The synthesis of 10 analogues of pepstatin modified so that statine is replaced by 4-amino-3-hydroxy-3,6-dimethylheptanoic acid (Me3Sta) or 4-amino-3-hydroxy-3-methyl-5-phenylpentanoic acid (Me3AHPPA) residues is reported. Both the 3S,4S and 3R,4S diastereomers of each analogue were tested as inhibitors of the aspartic proteases, porcine pepsin, cathepsin D, and penicillopepsin. In all cases ... More