Identification and partial characterization of factor Va heavy chain kinase from human platelets.

The Journal of Biological Chemistry
M Kalafatis

Abstract

Factor Va, the essential cofactor for prothrombinase, is phosphorylated on the acidic COOH terminus of the heavy chain of the cofactor, at Ser692, by a platelet membrane-associated casein kinase II (CKII). Consistent with this observation, phosphorylation of the factor Va heavy chain by the platelet kinase was inhibited by heparin. The membrane-associated platelet CKII kinase was partially purified using heparin-agarose, phosphocellulose, and ion exchange chromatography. CKII antigen was monitored using a polyclonal antibody to the alpha-subunit, and kinase activity in the various fractions was confirmed using human factor Va as a substrate. Immunoblotting experiments using polyclonal antibodies raised against synthetic peptides mimicking a portion of the deduced amino acid sequence of the alpha-, alpha'-, and beta-subunits of human CKII demonstrated the coexistence of both alpha- and alpha'-subunits in platelets and suggested that the platelet CKII kinase may exist in part as an alpha alpha'beta2 complex. It is also possible that there are two distinct populations of CKII in platelets, one that is alphaalpha/betabeta and one that is alpha'alpha'/betabeta. In the presence of the purified platelet-derived CKII, human factor Va i...Continue Reading

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Citations

Jul 14, 1999·Molecular Immunology·K Nilsson Ekdahl, B Nilsson
Feb 24, 2005·Current Opinion in Hematology·Michael Kalafatis
Sep 1, 2005·Journal of Thrombosis and Haemostasis : JTH·K Hansson, J Stenflo
Feb 24, 2017·Current Opinion in Chemical Biology·Bei ZhangHenry Daniell
Feb 24, 2007·Molecular & Cellular Proteomics : MCP·Geoff W BirrellMartin F Lavin
Sep 12, 2018·Platelets·Emmanuel AmpofoMichael D Menger
Apr 13, 2013·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Fabian StavenuiterMary J Heeb
Oct 24, 2002·Blood·Kenneth G Mann, Michael Kalafatis
Mar 13, 2003·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Flavio Meggio, Lorenzo A Pinna
Jun 6, 2003·The Journal of Biological Chemistry·Michael KalafatisKenneth G Mann
Jun 14, 2000·American Journal of Physiology. Heart and Circulatory Physiology·A BabinskaE Kornecki

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