The cDNA sequence and characterization of the Ca2+/calmodulin-dependent protein kinase-Gr from human brain and thymus.
We have isolated and sequenced cDNAs encoding Ca2+/calmodulin-dependent protein kinase type Gr (CaM-K-Gr, also called CaM-K-IV) from human brain and thymus. The sequence of the protein coding region of the cDNA is identical in both brain and thymus, although Northern hybridization analysis shows variation of the mRNA transcripts in these tissues. The sequence predicts a protein of M(r) 51,897 that is 83.7% identical and shows 89.2% similarity with the rat homologue. The deduced human CaM-K-Gr is identical to the rat and mouse proteins in the portion of the enzyme involved in ATP binding, the catalytic domain and Ca2+/calmodulin-binding domain; however, the N terminus of the human kinase, which may comprise a second regulatory domain [McDonald et al., J. Biol. Chem. 268 (1993) 10054-10059], contains a 4-amino-acid (aa) insertion relative to the rodent enzymes. Additionally, the C-terminal association domain shows only 45.2 and 41.6% identity with the rat and mouse proteins, respectively, suggesting that this domain is not constrained by stringent structural and functional requirements. Based on the predicted aa sequence of the human kinase, we produced polyclonal antisera against a C-terminal peptide that recognizes two forms of CaM-K-Gr in human T-cell lymphoma and neuroblastoma cell lines. The human antiserum cross-reacts with the rat and mouse proteins and immunoprecipitates the active kinase.
Bland
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Wellcome Research Laboratories, Research Triangle Park, NC 27709.
Monroe
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Ohmstede
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D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D001483:Base Sequence; D054734:Calcium-Calmodulin-Dependent Protein Kinase Type 4; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D002531:Cerebellum / Q000737:chemistry* / Q000201:enzymology; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010766:Phosphorylation; D012333:RNA, Messenger / Q000032:analysis; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013737:Testis / Q000737:chemistry; D013950:Thymus Gland / Q000737:chemistry* / Q000201:enzymology; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D000906:Antibodies; D018076:DNA, Complementary; D012333:RNA, Messenger; C517683:CAMK4 protein, human; D054734:Calcium-Calmodulin-Dependent Protein Kinase Type 4; D017871:Calcium-Calmodulin-Dependent Protein Kinases; C517685:Camk4 protein, mouse; C517686:Camk4 protein, rat