Endogenous ADP-ribosylation in human platelets.
We have studied ADP-ribosyltransferase activity in platelet cytosol and electropermeabilized platelets. Cytosolic activity causes ADP-ribosylation or of a 37 kDa protein that is activated by increasing the concentration of potassium phosphate. ADP-ribosylation is inhibited by thiol reagents, an effect partially reversed by cholera toxin. In electropermeabilized platelets incubated with [alpha-32P]NAD, the 37 kDa protein is also ADP-ribosylated as are other proteins and albumin. Under these conditions, ADP-ribosylation is partially inhibited by nicotinamide. This experimental design could be used to determine the effect of cell agonists on endogenous ADP-ribosylation of proteins.
Molina y Vedia
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L
Division of Cell Biology, Burroughs Wellcome Co., Research Triangle Park, NC 27709.
Nolan
R D
RD
Lapetina
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EG
D000246:Adenosine Diphosphate Ribose / Q000097:blood*; D001792:Blood Platelets / Q000201:enzymology*; D002772:Cholera Toxin / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D003600:Cytosol / Q000201:enzymology; D004229:Dithiothreitol / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D009243:NAD / Q000378:metabolism; D037342:Pertussis Toxin; D010710:Phosphates / Q000494:pharmacology; D011065:Poly(ADP-ribose) Polymerases / Q000097:blood*; D011188:Potassium / Q000494:pharmacology; D017680:Potassium Compounds; D012709:Serum Albumin / Q000378:metabolism; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D010710:Phosphates; D017680:Potassium Compounds; D012709:Serum Albumin; D010566:Virulence Factors, Bordetella; D009243:NAD; D000246:Adenosine Diphosphate Ribose; D002772:Cholera Toxin; C013216:potassium phosphate; D011065:Poly(ADP-ribose) Polymerases; D037342:Pertussis Toxin; D008274:Magnesium; D011188:Potassium; D004229:Dithiothreitol