Open AccessMolecular cloning, characterization, and expression of a cDNA encoding the "80- to 87-kDa" myristoylated alanine-rich C kinase substrate: a major cellular substrate for protein kinase C.
Author(s) -
Deborah J. Stumpo,
Jonathan M. Graff,
Katherine A. Albert,
Paul Greengard,
Perry J. Blackshear
Publication year - 1989
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.86.11.4012
Subject(s) - complementary dna , microbiology and biotechnology , marcks , myristoylation , open reading frame , biology , protein kinase a , amino acid , protein kinase c , peptide sequence , biochemistry , protein a/g , alanine , kinase , gene , phosphorylation , fusion protein , recombinant dna
We isolated and sequenced a cDNA clone encoding the bovine "80- to 87-kDa" protein, a major cellular substrate for protein kinase C. An open reading frame of 1005 base pairs predicted a protein of 335 amino acids (Mr, 31,949). Despite this predicted size, the protein migrated on SDS/polyacrylamide gels with an apparent molecular weight of 80-87,000 after expression of the cDNA in cells lacking the protein. It was highly enriched in alanine (28.4 mol %), contained an amino-terminal myristoylation consensus sequence, and included a 25-residue basic domain containing the known protein kinase C phosphorylation sites. Two mRNA species (2.6 and 4.4 kilobases) were most highly expressed in brain, spinal cord, spleen, and lung, in parallel with the distribution of immuno-reactive protein. Genomic blot analysis indicated the likelihood of a single gene coding for this mRNA. We propose the name myristoylated alanine-rich C kinase substrate (MARCKS) for this protein.