Inhibition of NADPH oxidase activation by 4-(2-aminoethyl)-benzenesulfonyl fluoride and related compounds.
AuthorsDiatchuk V, Lotan O, Koshkin V, Wikstroem P, Pick E
JournalJ Biol Chem
PubMed ID9148950
The elicitation of an oxidative burst in phagocytes rests on the assembly of a multicomponental complex (NADPH oxidase) consisting of a membrane-associated flavocytochrome (cytochrome b559), representing the redox element responsible for the NADPH-dependent reduction of oxygen to superoxide (O-2), two cytosolic components (p47(phox), p67(phox)), and the small GTPase Rac (1 ... 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
Proteolytic processing of Alzheimer's disease beta A4 amyloid precursor protein in human platelets.
AuthorsLi QX, Evin G, Small DH, Multhaup G, Beyreuther K, Masters CL
JournalJ Biol Chem
PubMed ID7775475
'The processing of amyloid precursor protein (APP) and production of beta A4 amyloid are events likely to influence the development and progression of Alzheimer''s disease, since beta A4 is the major constituent of amyloid deposited in this disorder. Our previous studies showed that human platelets contain full-length APP (APPFL) 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
Lysine- and arginine-specific proteinases from Porphyromonas gingivalis. Isolation, characterization, and evidence for the existence of complexes with hemagglutinins.
AuthorsPike R, McGraw W, Potempa J, Travis J
JournalJ Biol Chem
PubMed ID8276827
'Porphyromonas gingivalis contains many virulence factors that have been implicated as participants in the progression of periodontal disease. It has been shown to produce proteinases of "trypsin-like" specificity in a number of molecular forms, but previous work in our laboratory resulted in the purification of a major arginine-specific cysteine proteinase, ... 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
Ca2+ ionophore A23187 and thrombin inhibit the pertussis-toxin-induced ADP-ribosylation of the alpha-subunit of the inhibitory guanine-nucleotide-binding protein and other proteins in human platelets.
AuthorsSiess W, Wilhelm B, Aepfelbacher M, Gennity JM
JournalEur J Biochem
PubMed ID1935970
'Inhibitory guanine-nucleotide-binding proteins (Gi proteins) are substrates for pertussis toxin and the decreased pertussis-toxin-dependent ADP ribosylation of Gi proteins upon prior specific hormonal stimulation of cells is thought to reflect the receptor-mediated activation of Gi proteins, leading to their subsequent dissociation into alpha i and beta/gamma subunits. In the present ... More
Calcium-activated DNA fragmentation kills immature thymocytes.
AuthorsMcConkey DJ, Hartzell P, Nicotera P, Orrenius S
JournalFASEB J
PubMed ID2497041
'Glucocorticoid hormones kill immature thymocytes by activating a self-destructive process that involves extensive DNA fragmentation. It has been demonstrated that thymocyte suicide is dependent on an early, sustained increase in cytosolic Ca2+ concentration, and new protein synthesis, but the biochemical lesion that leads to cell death has not been established. ... 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
Glucose, other secretagogues, and nerve growth factor stimulate mitogen-activated protein kinase in the insulin-secreting beta-cell line, INS-1.
AuthorsFrödin M, Sekine N, Roche E, Filloux C, Prentki M, Wollheim CB, Van Obberghen E
JournalJ Biol Chem
PubMed ID7713882
'The signaling pathways whereby glucose and hormonal secretagogues regulate insulin-secretory function, gene transcription, and proliferation of pancreatic beta-cells are not well defined. We show that in the glucose-responsive beta-cell line INS-1, major secretagogue-stimulated signaling pathways converge to activate 44-kDa mitogen-activated protein (MAP) kinase. Thus, glucose-induced insulin secretion was found to ... 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
Isolation and characterization of leupeptins produced by Actinomycetes.
AuthorsKondo SI, Kawamura K, Iwanaga J, Hamada M, Aoyagi T
JournalChem Pharm Bull (Tokyo)
PubMed ID5347594
Tackling the protease problem in Saccharomyces cerevisiae.
AuthorsJones EW
JournalMethods Enzymol
PubMed ID2005802
A neutral thiol protease secreted from newly excysted metacercariae of trematode parasite Paragonimus westermani: purification and characterization.
AuthorsYamakami K, Hamajima F
JournalComp Biochem Physiol B
PubMed ID2344732
1. A neutral thiol protease was purified from the culture filtrate of newly excysted metacercariae of Paragonimus westermani to homogeneity as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, having a monomeric form with mol. wt 22,000. 2. It expressed activity on t-butyloxycarbonyl-valyl-leucyl-lysyl-4-methyl-coumaryl-7-amide in the presence of cysteine at an optimal ... More
Memory formation in the chick depends on membrane-bound protein kinase C.
AuthorsBurchuladze R, Potter J, Rose SP
JournalBrain Res
PubMed ID2292019
The role of protein kinase C (PKC) in the formation of memory for a one-trial passive avoidance task in 1-day-old chicks has been studied, following earlier observations that training on this task results in transient and lateralised changes in the phosphorylation state of presynaptic B-50 protein, a PKC substrate. In ... More
Phorbol ester-induced morphological changes in transformed chick fibroblasts: evidence for direct catalytic involvement of plasminogen activator.
AuthorsQuigley JP
JournalCell
PubMed ID222474
The tumor promoter phorbol myristate acetate (PMA) induces the production of the serine protease plasminogen activator (PA) in cultures of normal chick embryo fibroblasts (CEF) and synergistically enhances PA production in Rous sarcoma virus-transformed chick embryo fibroblasts (RSVCEF). Following PMA treatment of serum-free RSVCEF cultures, PA induction is accompanied by ... 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
Purification and characterization of UDP-glucose:ceramide glucosyltransferase from rat liver Golgi membranes.
AuthorsPaul P, Kamisaka Y, Marks DL, Pagano RE
JournalJ Biol Chem
PubMed ID8567691
We present a method for solubilizing and purifying UDP-Glc:ceramide glucosyltransferase (EC 2.4.1.80; glucosylceramide synthase (GCS) from a rat liver and present data on its substrate specificity. A Golgi membrane fraction was isolated, washed with N-lauroylsarcosine, and subsequently treated with 3[3-cholamidopropyl)-dimethylammonio]-2-hydroxy-1-propanesulfonate to solubilize the enzyme. GCS activity was monitored throughout purification ... 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
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
GABAA/benzodiazepine receptors in acutely isolated hippocampal astrocytes.
The properties of GABA receptor-mediated responses were examined in noncultured astrocytes, acutely isolated from the mature rat hippocampus. Whole-cell patch clamping revealed a GABA-activated Cl- conductance that was mimicked by the GABAA receptor agonist muscimol and depressed by the GABAA antagonists bicuculline and picrotoxin. The GABAA-activated currents were potentiated by ... More
Activity-induced internalization and rapid degradation of sodium channels in cultured fetal neurons.
A regulatory mechanism for neuronal excitability consists in controlling sodium channel density at the plasma membrane. In cultured fetal neurons, activation of sodium channels by neurotoxins, e.g., veratridine and alpha-scorpion toxin (alpha-ScTx) that enhance the channel open state probability induced a rapid down-regulation of surface channels. Evidence that the initial ... 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
Coordinated regulation of platelet actin filament barbed ends by gelsolin and capping protein.
Exposure of cryptic actin filament fast growing ends (barbed ends) initiates actin polymerization in stimulated human and mouse platelets. Gelsolin amplifies platelet actin assembly by severing F-actin and increasing the number of barbed ends. Actin filaments in stimulated platelets from transgenic gelsolin-null mice elongate their actin without severing. F-actin barbed ... More