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PMID: 2470091 Published · ppublish English Journal Article

A ras-related protein is phosphorylated and translocated by agonists that increase cAMP levels in human platelets.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 86 ·No. 9 ·1989-05-00 ·Pages 3131-4

Lapetina EG, Lacal JC, Reep BR, Molina y Vedia L

Abstract

The antigenicity of platelet proteins was assayed against various monoclonal antibodies (mAbs) that recognize specific epitopes of the ras-encoded p21 protein. mAb M90, which detects the region of p21 protein within amino acids 107-130 and inhibits its GTP-binding activity, strongly reacted with a 22-kDa protein present in the particulate fraction of human platelets. Other mAbs against ras-encoded proteins, including Y13-259, which efficiently detects ras proteins from a variety of organisms, did not recognize the platelet 22-kDa protein. Transfer of the platelet 22-kDa protein to nitrocellulose paper showed that the protein binds [alpha-32P]GTP. Moreover, preincubation of the transferred protein with mAb M90 drastically reduced its GTP-binding activity. Treatment of platelets with iloprost, a prostacyclin analog, caused (i) a time-dependent increase of a 24-kDa protein that is recognized by mAb M90 in particulate and cytosolic fractions and (ii) the gradual decrease of the 22-kDa protein from the particulate fraction. When platelets were labeled with 32P and then treated with iloprost, the 24-kDa protein was found to be phosphorylated. The 32P-labeled 24-kDa protein was specifically immunoprecipitated by mAb M90. These results suggest that appearance of the 24-kDa protein results from phosphorylation of the 22-kDa protein, which shifts its mobility to a higher molecular mass area.

MeSH Terms
Antibodies, Monoclonal Antibody Specificity Binding Sites Biological Transport Blood Platelets/drug effects,metabolism Cell Membrane/metabolism Cyclic AMP/blood Electrophoresis, Polyacrylamide Gel Epitopes/immunology Epoprostenol/pharmacology Guanosine Triphosphate/blood Humans Iloprost Immunosorbent Techniques Membrane Proteins Molecular Weight Phosphorylation Protein Kinases/blood Proto-Oncogene Proteins/blood Proto-Oncogene Proteins p21(ras)
Chemicals
Antibodies, Monoclonal Epitopes Membrane Proteins Proto-Oncogene Proteins Guanosine Triphosphate Epoprostenol Cyclic AMP Protein Kinases HRAS protein, human Proto-Oncogene Proteins p21(ras) Iloprost
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Lapetina E G
Division of Cell Biology, Burroughs Wellcome Company, Research Triangle Park, NC 27709.
Lacal J C
Reep B R
Molina y Vedia L
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.86.9.3131
Published
1989-05-00
Pages
3131-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC287079
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.86.9.3131 PMC Full Text View journal details
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