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Multisite phosphorylation of the 80 kDa (MARCKS) protein kinase C substrate in C3H/10T1/2 fibroblasts Quantitative analysis of individual sites by solid‐phase microsequencing
Author(s) -
Arness Bob,
Manjarrez-Hernandez H.Angel,
Howell Steve A.,
Learmonth Michele,
Aitken Alastair
Publication year - 1992
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(92)80557-w
Subject(s) - phosphorylation , okadaic acid , protein kinase c , protein phosphorylation , kinase , peptide , biochemistry , marcks , protein kinase a , chemistry , in vitro , substrate level phosphorylation , microbiology and biotechnology , enzyme , phosphatase , biology
A synthetic peptide, KKKKRFSFKKSFKLSGFSFKK, containing the phosphorylation sites of the acidic 80–87 kDa protein kinase C substrate was used to identify phosphopeptides in enzyme digests of this protein from mouse fibroblast C3H/10T1/2 cells. Stimulation of phosphorylation occurred, in vivo, with TPA at Ser 7 , Ser 11 and Ser 18 , and, will two less potent phorbol esters, at Ser 7 and Ser 18 . Okadaic acid effected a net phosphorylation of Ser 7 and/or Ser 11 . Solid‐phase sequencing showed that, in vitro, the order of initial rate of phosphorylation was Ser 11 > Ser 7 > Ser 18 , while Ser 18 was preferentially phosphorylated when either Ser 7 or Ser 11 was occupied. No significant phosphorylation of Ser 15 was detected.