Isolation and Characterization of a Mouse Protein C cDNA

• Published in: Journal of biochemistry (Tokyo) Vol. 111; no. 4; pp. 491 - 495
• Main Authors: Tada, Norihiro, Sato, Masahiro, Tsujimura, Atsushi, Iwase, Reiko, Hashimoto-Gotoh, Tamotsu
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 01.04.1992
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Biological and medical sciences; Blotting, Northern; cDNA; Cloning, Molecular; DNA - analysis; DNA - isolation & purification; Fundamental and applied biological sciences. Psychology; genes; Genes. Genome; Humans; liver; Mice; Mice, Inbred BALB C; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; nucleotide sequence; predictions; protein C; Protein C - genetics; Sequence Homology, Nucleic Acid; Northern blotting; Vertebrata; Mammalia; Complementary DNA; Mouse; Active site; Rodentia; Protein C; Primary structure; Binding site; Gene expression
• ISSN: 0021-924X; 1756-2651
• DOI: 10.1093/oxfordjournals.jbchem.a123785

Protein C (PC) i8 a vitamin K-dependent serine protease, a deficiency of which results In thrombus. There Is no spontaneously occurring mouse model of the disease. Attempts to create such a model in mice by using anti-sense gene technology requires Isolation of a normal mouse PC cDNA. When a mouse liver (BALB/c) cDNA library was screened using a human PC cDNA as a probe, nine overlapping cDNA clones were isolated and sequenced. The cloned mouse PC cDNA comprised 1,512 nucleotldes and the open reading frame of the cDNA encoded a polypeptide of 4431 amino acids residues including a leader peptide composed of 41 amino acids. Mouse PC exhibited high homology to both human and bovine PCs. Mouse PC also had several structural features common in other PCs; locations of 23 Cys residues, location of putative β-hydroxyAsp71, possible carbohydrate attachment sites involving Asp residues at amino acid positions 249,314, and 330, and location of active sites such as His212, Asp258, and Ser361. Northern blot hybridization analysis identified a single species of mouse PC mRNA (2.0 kb in length) In mouse liver.