Expression of the cDNA encoding lipocortin-like 39 kDa protein of guinea pig neutrophils in yeast: Purification and biological characterization

• Published in: FEBS letters 1989-09, Vol.255 (2), p.231-236
• Main Authors: Sato, Eisuke F., Tanaka, Yoshikazu, Edashige, Keisuke, Kobuchi, Hirotsugu, Morishita, Seiji, Sugino, Yasuo M., Inoue, Masayasu, Utsumi, Kozo
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Annexins; Binding and carrier proteins; Biological and medical sciences; Calcium-Binding Proteins - blood; Calcium-Binding Proteins - genetics; Calcium-Binding Proteins - isolation & purification; Cloning, Molecular; DNA - genetics; Fundamental and applied biological sciences. Psychology; Genetic Vectors; Guinea Pigs; Lipocortin; Molecular Weight; Neutrophil; Neutrophils - metabolism; Phospholipases - antagonists & inhibitors; Plasmids; Protein kinase; Proteins; Recombinant protein; Recombinant Proteins - isolation & purification; Saccharomyces cerevisiae - genetics
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(89)81097-X

The cDNA encoding lipocortin-like 39 kDa protein in guinea pig neutrophils was cloned into a yeast expression vector and the constructed plasmid was introduced into a yeast. The gene was expressed in an eukaryotic cell, yeast Saccharomyces cerevisiae and the recombinant protein was purified and characterized. The purified protein was identical with the native one with respect to the antigenicity and several biochemical properties, such as inhibitory action against phospholipase A 2, Ca 2+-dependent binding to acidic-phospholipids and F-actin and availability as a substrate for tyrosine kinase (EGF receptor/kinase) and protein kinase C.