Primary structure of bovine calpactin I heavy chain (p36), a major cellular substrate for retroviral protein-tyrosine kinases: homology with the human phospholipase A2 inhibitor lipocortin

• Published in: Biochemistry (Easton) Vol. 25; no. 16; pp. 4497 - 4503
• Main Authors: KRISTENSEN, T, SARIS, C. J. M, HUNTER, T, HICKS, L. J, NOONAN, D. J, GLENNEY, J. R. JR, TACK, B. F
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 12.08.1986
• Subjects: Amino Acid Sequence; Analytical, structural and metabolic biochemistry; Animals; Annexins; Base Sequence; Binding and carrier proteins; Biological and medical sciences; Calcium-Binding Proteins - genetics; Calcium-Binding Proteins - isolation & purification; Calcium-Binding Proteins - metabolism; Cattle; Cell Line; DNA - metabolism; Fundamental and applied biological sciences. Psychology; Glycoproteins - genetics; Humans; Macromolecular Substances; Peptide Fragments - analysis; Phospholipases A - antagonists & inhibitors; Phospholipases A2; Plasmids; Protein-Tyrosine Kinases - metabolism; Proteins; Sequence Homology, Nucleic Acid; Trypsin; Calcium; heavy-Peptide chain; Ox; Homology; Gene organization; Primary structure; Substrate; Binding protein; Vertebrata; Mammalia; Multigene family; Artiodactyla; Protein-tyrosine kinase; Ungulata
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi00364a007

An amplified Okayama-Berg plasmid cDNA library was constructed from total poly(A)+ RNA isolated from the Madin-Darby bovine kidney cell line MDBK. This library was screened with a partial murine calpactin I heavy chain (p36) cDNA clone, the identification of which was based on bovine p36 tryptic peptide sequences generated during the course of these studies. The largest p36 cDNA insert (p36/6 of 1.6 kilobase pairs) was fully sequenced by the dideoxy method. The DNA sequence of this insert had an open reading frame of 1014 base pairs and coded for a protein with a molecular weight of 38 481. The deduced protein sequence of 338 residues was concordant with 173 residue positions of p36 determined at the protein level. The 5'- and 3'-ends of p36/6 contained 54 and 307 base pairs of untranslated sequence, respectively. Examination of poly(A)+ RNA prepared from the Madin-Darby cell line indicated a p36 mRNA species of about 1.6 kilobases. Four regions of internal homology, each about 70 amino acid residues in length, were observed in the deduced protein sequence for p36. Thirty-three of the 70 residue positions were conserved in at least three of the four repeating units. A comparison of derived amino acid sequence for bovine p36 with that previously determined for human lipocortin [Wallner, B. P., Mattaliano, R. J., Hession, C., Cate, R. L., Tizard, R., Sinclair, L. K., Foeller, C., Chow, E. P., Browning, J. L., Ramachandran, K. L., & Pepinsky, R. B. (1986) Nature (London) 320, 77-81] revealed extensive homology (66% overall) and the presence of four repetitive regions in the lipocortin structure.