Site‐directed mutation makes rabbit calcyclin dimer

FEBS Letters

Akatsuka, Hajime¹  Tokumitsu, Hiroshi¹  Hidaka, Hiroyoshi¹  Ando, Yuhko¹  Watanabe, Masato¹ 

[1] Department of Pharmacology, Nagoya University School of Medicine, Showa-ku, Nagoya 466, Japan

关键词: EF-hand protein;    Calcyclin;    cDNA cloning;    Expression of cDNA;    SDS;    sodium dodecyl sulfate;    PAGE;    polyacrylamide gel electrophoresis;    bp;    base pairs;    PCR;    polymerase chain reaction;    BPB;    bromophenol blue;    Tricine;    Tris(hydroxymethyl)methylglycine;    IPTG;    isopropyl-β-d-thiogalactoside;   

DOI  :  10.1016/0014-5793(92)80953-E

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Unlike human, rat and mouse calcyclin, purified rabbit calcyclin did not form a dimer on Tricine SDS-PAGE under non-reduced conditions. Based on the internal peptide sequence of rabbit calcyclin, we isolated and sequenced a cDNA clone encoding calcyclin. The sequence of this clone (pCaiC) is 629 bp long and codes 90 amino acid residues of a protein with a molecular mass of 10,153 Da. By Northern blot analysis, a major band of 0.9 kbp and a minor band of 2.6 kbp were detected in the lung. The recombinant calcyclin mutated serine at the third position to cysteine was expressed in E. coli and made dimer formation under non-reduced conditions on SDS-PAGE. Whether or not this type of mutation which prevents dimer formation of calcyclin plays a physiological role in the rabbit lung is the subject of an ongoing study.

【 授权许可】

Unknown