Purification and partial characterization of a calmodulin‐like protein from cell suspension cultures of Catharanthus roseus

FEBS Letters

DiCosmo, Frank¹  Radvanyi, Laszlo G.¹ 

[1] Centre for Plant Biotechnology, Department of Botany, University of Toronto Ontario M5S 1A1, Canada

关键词: Calmodulin;    Phosphodiesterase;    Enzyme activator;    Protein purification;    Cell suspension;    (Cantharanthus roseus);    K 1;    half-inhibition constant;    SDS-PAGE;    SDS-polyacrylamide gel electrophoresis;    PMSF;    phenylmethylsulphonyl fluoride;    PVPP;    polyvinylpolypyrollidone;    nkat;    nanokatal;    PDE;    phosphodiesterase;    HPLC;    high-performance liquid chromatography;   

DOI  :  10.1016/0014-5793(88)80131-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A calmodulin-like protein was isolated from suspension cultured cells of Catharanthus roseus and purified by a combination of Ca²⁺-facilitated phenyl-Sepharose affinity chromatography and reverse-phase HPLC. The HPLC-purified protein was analysed using SDS-PAGE and found to be a homogeneous 19.5 kDa band in gels containing 1 mM EGTA unlike a higher plant calmodulin from spinach which migrated as a 17.38 kDa band. Despite this apparent difference in molecular mass, the purified protein showed a similar increase in electrophoretic mobility (4010 Da) to spinach calmodulin (4200 Da) in gels containing 1 mM Ca²⁺ and had a plant calmodulin-like UV spectrum and phosphodiesterase-activation profile.

【 授权许可】

Unknown