Rabbit fast skeletal muscle phospholipase C Molecular weight determination by renaturation after polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate

FEBS Letters

Heilmeyer, Ludwig M.G.¹  Windhofer, Volker¹  Varsànyi, Magdolna¹ 

[1] Institut für Physiologische Chemie, Abteilung für Biochemie Supramolekularer Systeme, Ruhr-Universität Bochum, Universitätsstr. 150, 4630 Bochum I, Germany

关键词: Phospholipase C;    Fast skeletal muscle;    Molecular weight determination;    Renaturation;    Ins(1;    4;    5)P3;    d-myo-inositol 1;    4;    5-triphosphate;    PtdIns;    1-(3-sn-phosphatidyl)-1d-myo-inositol;    PtdIns(4)P;    1-(3-sn-phosphatidyl)-1d-myo-inositol-4-phospahte;    PtdIns(4;    5)P2;    1-(3-sn-phosphatidyl)-1d-myo-inositol-4;    5-bisphosphate;    PtdSer;    (3-sn-phosphatidyl)2;    serine;    PtdFln;    (3-sn-phosphatidyl)ethanolamine;    DOC;    deoxycholate;    SDS;    sodium dodecylsulfate;   

DOI  :  10.1016/0014-5793(92)81182-L

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Phosphoinositide specific phospholipase C from rabbit fast skeletal muscle has been enriched ca. 1,000-fold with a specific activity of 40 μmol × min⁻¹ × mg⁻¹. Following SDS-PAGE, renaturation of the enzyme protein in the presence of deoxycholate allowed the determination of an apparent molecular weight of 110 kDa. Gel-filtration of the native enzyme resulted in a very similar apparent molecular weight of 115 kDa, however, associated proteins of higher molecular weight were also found. Free Ca²⁺ concentrations needed for half-maximal activation of PtdIns(4,5)P₂, PtdIns4P and PtdIns hydrolysis are 6.3 μM, 85 μM and 1.8 mM, and the K_m values for these substrates 102, 340 and 937 μM, respectively.

【 授权许可】

Unknown