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Predicate | Object |
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rdf:type | |
lifeskim:mentions |
umls-concept:C0026385,
umls-concept:C0027950,
umls-concept:C0068355,
umls-concept:C0086376,
umls-concept:C0205251,
umls-concept:C0752214,
umls-concept:C0967718,
umls-concept:C1383501,
umls-concept:C1514873,
umls-concept:C1546857,
umls-concept:C1556066,
umls-concept:C1619636,
umls-concept:C1879547,
umls-concept:C2699488
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pubmed:issue |
21
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pubmed:dateCreated |
1991-8-23
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pubmed:abstractText |
Activation of the membrane-associated NADPH oxidase in intact human neutrophils requires a receptor-associated heterotrimeric GTP-binding protein that is sensitive to pertussis toxin. Activation of this NADPH oxidase by arachidonate in a cell-free system requires an additional downstream pertussis toxin-insensitive G protein (Gabig, T. G., English, D., Akard, L. P., and Schell, M. J. (1987) (J. Biol. Chem. 262, 1685-1690) that is located in the cytosolic fraction of unstimulated cells (Gabig, T. G., Eklund, E. A., Potter, G. B., and Dykes, J. R. (1990) J. Immunol. 145, 945-951). In the present study, immunodepletion of G proteins from the cytosolic fraction of unstimulated neutrophils resulted in a loss of the ability to activate NADPH oxidase in the membrane fraction. The activity in immunodepleted cytosol was fully reconstituted by a partially purified fraction from neutrophil cytosol that contained a 21-kDa GTP-binding protein. Purified human recombinant Krev-1 p21 also completely reconstituted immunodepleted cytosol whereas recombinant human H-ras p21 or yeast RAS GTP-binding proteins had no reconstitutive activity. Rabbit antisera raised against a synthetic peptide corresponding to the effector region of Krev-1 (amino acids 31-43) completely inhibited cell-free NADPH oxidase activation, and this inhibition was blocked by the synthetic 31-43 peptide. An inhibitory monoclonal antibody specific for ras p21 amino acids 60-77 (Y13-259) had no effect on cell-free NADPH oxidase activation. Activation of the NADPH oxidase in intact neutrophils by stimulation with phorbol myristate acetate caused a marked increase in the amount of membrane-associated antigen recognized by 151 antiserum on Western blot. Thus a G protein in the cytosol of unstimulated neutrophils antigenically and functionally related to Krev-1 may be the downstream effector G protein for NADPH oxidase activation. This system represents a unique model to study molecular interactions of a ras-like G protein.
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pubmed:grant | |
pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/GTP-Binding Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/NADH, NADPH Oxidoreductases,
http://linkedlifedata.com/resource/pubmed/chemical/NADPH Oxidase,
http://linkedlifedata.com/resource/pubmed/chemical/Oligonucleotides,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/rap GTP-Binding Proteins
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pubmed:status |
MEDLINE
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pubmed:month |
Jul
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pubmed:issn |
0021-9258
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
25
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pubmed:volume |
266
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
13964-70
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pubmed:dateRevised |
2007-11-14
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pubmed:meshHeading |
pubmed-meshheading:1906890-Cell-Free System,
pubmed-meshheading:1906890-Cytosol,
pubmed-meshheading:1906890-Enzyme Activation,
pubmed-meshheading:1906890-GTP-Binding Proteins,
pubmed-meshheading:1906890-Humans,
pubmed-meshheading:1906890-NADH, NADPH Oxidoreductases,
pubmed-meshheading:1906890-NADPH Oxidase,
pubmed-meshheading:1906890-Neutrophils,
pubmed-meshheading:1906890-Oligonucleotides,
pubmed-meshheading:1906890-Polymerase Chain Reaction,
pubmed-meshheading:1906890-Recombinant Proteins,
pubmed-meshheading:1906890-Signal Transduction,
pubmed-meshheading:1906890-Structure-Activity Relationship,
pubmed-meshheading:1906890-rap GTP-Binding Proteins
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pubmed:year |
1991
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pubmed:articleTitle |
Resolution of a low molecular weight G protein in neutrophil cytosol required for NADPH oxidase activation and reconstitution by recombinant Krev-1 protein.
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pubmed:affiliation |
Department of Medicine, Indiana University School of Medicine, Indianapolis 46202.
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pubmed:publicationType |
Journal Article,
In Vitro,
Research Support, U.S. Gov't, P.H.S.
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