Substrates for the cell surface protein kinase in blood: a calf serum protein and its precursor in plasma.
A protein in calf serum with molecular mass of 125,000 is selectively phosphorylated by the surface kinase activity of intact tissue culture cells and erythrocytes. The protein, termed pp125, is phosphorylated at serine and threonine residues to a ratio of greater than 1 mol P/mol. The pp125 is an acidic protein (pI 4.4) which also serves as substrate for purified phosvitin/casein kinases but not for cyclic AMP-dependent protein kinases. About 80-fold purification of pp125 was achieved by ion exchange and affinity chromatography. Gel filtration under non-reducing conditions showed that pp125 is part of a complex (Mr 535,000). The pp125 obviously originates from a large plasma protein: the incubation of calf plasma with intact cells in the presence of [gamma-32P]ATP resulted in the labeling of a protein with Mr greater than 300,000 (pp greater than 300). The relationship between pp greater than 300 in plasma and pp125 in serum was demonstrated by cyanogen bromide peptide patterns, and the use of specific anti-serum raised against pp125. Furthermore, it was shown that pp125 is derived from pp greater than 300 during blood clotting.
Institute of Experimental Pathology, German Cancer Research Center, Heidelberg, Federal Republic of Germany.
D
D
Kübler
M
M
Fehst
T
T
Garcon
W
W
Pyerin
E
E
Burow
V
V
Kinzel
D000818:Animals; D001798:Blood Proteins / Q000302:isolation & purification / Q000378:metabolism; D002417:Cattle; D002462:Cell Membrane / Q000201:enzymology*; D004910:Erythrocyte Membrane / Q000201:enzymology*; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D007106:Immune Sera; D007525:Isoelectric Focusing; D007700:Kinetics; D008970:Molecular Weight; D010449:Peptide Mapping; D010766:Phosphorylation; D011494:Protein Kinases / Q000097:blood / Q000378:metabolism*
D001798:Blood Proteins; D007106:Immune Sera; D011494:Protein Kinases