Molecular cloning and predicted full‐length amino acid sequence of the type Iβ isozyme of cGMP‐dependent protein kinase from human placenta

FEBS Letters

Lohmann, Suzanne M.¹  Kalderon, Daniel²  Ronander, Inger³  Jahnsen, Tore³  Natarajan, Vasanti³  Sandberg, Mårten³  Walter, Ulrich¹ 

[1] Medizinische Universitätsklinik, Labor für Klinische Biochemie, D-8700 Würzburg, FRG;Department of Biological Sciences, Columbia University, New York, NY 10072, USA;Laboratory for Gene Technology, Institute of Pathology, Rikshospitalet, University of Oslo, N-0027 Oslo 1, Norway

关键词: Protein kinase;    cGMP-dependent;    Molecular cloning;    cDNA sequence;    Tissue specific expression;    (Human placenta);   

DOI  :  10.1016/0014-5793(89)81114-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

In this study we report the isolation and characterization of three overlapping cDNA clones for the type Iβ isozyme of cGMP-dependent protein kinase (cGK) from human placenta libraries. The composite sequence was 3740 nucleotides long and contained 58 nucleotides from the 5′-noncoding region, an open reading frame of 2061 bases including the stop codon, and a 3′-noncoding region of 1621 nucleotides. The predicted full-length human type Iβ cGK protein contained 686 amino acids including the initiator methionine, and had an estimated molecular mass of 77 803 Da. On comparison to the published amino acid sequence of bovine lung Iα, human placenta Iβ cGK differed by only two amino acids in the carboxyl-terminal region (amino acids 105–686). In contrast, the amino-terminal region of the two proteins was markedly different (only 36% similarity), and human Iβ cGK was 16 amino acids longer. In a specific region in the amino-terminus (amino acids 63–75), 12 out of 13 amino acids of the human Iβ cGK were identical to the partial amino acid sequence recently published for a new Iβ isoform of cGK from bovine aorta. Northern blot analysis demonstrated a human Iβ cGK mRNA, 7 kb in size, in human uterus and weakly in placenta. An mRNA of 7 kb was also observed in rat cerebellum, cerebrum, lung, kidney, and adrenal, whereas an mRNA doublet of 7.5 and 6.5 kb were observed in rat heart. Comparison of Northern and Western blot analyses demonstrated that the mRNA and protein for cerebellar cGK increased during the development of rats from 5 to 30 days old, whereas the 6.5 kb mRNA in rat heart declined.

【 授权许可】

Unknown