A cross‐linked complex between horse pancreatic lipase and colipase

FEBS Letters

Chapus, Catherine¹  Chaillan, Catherine¹  Lombardo, Dominique¹  Rogalska, Ewa¹ 

[1] CNRS, Centre de Biochimie et de Biologie Moléculaire, 31 Chemin Joseph-Aiguier, BP 71, 13402 Marseille Cedex 9, France

关键词: Lipase;    Colipase;    Complex;    Crosslinking;    CME-CDI;    N-cyclohexyl-N'-2-morpholinoethyl-carbodiimide-methyl-p-toluoisulfonate;    NaTDC;    sodium taurodeox-ycholate;    BCIP;    5-bromo-4-chloro-3-indolyl phosphate;   

DOI  :  10.1016/0014-5793(89)81592-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The water soluble carbodiimide N-cyclohexyl-N'-2-morpholinoethyl-carbodiimide-methyl-p-toluolsulfonate was found to effectively covalently cross-link pancreatic colipase to lipase as evidenced by Western blotting experiments using antibodies directed either against lipase or colipase. Moreover the resulting covalent complex has a M _r, consistent with a stoichiometry of 1 mol colipase per mol lipase. Cross-linked lipase and colipase retain their activity implying a correct covalent binding between the two proteins. The specificity of the lipase-colipase binding was further supported by the very low amount of cross-linked products when lipase or colipase alone were incubated in the presence of carbodiimide. The formation of a covalent lipase-colipase complex in the presence of carbodiimide clearly demonstrates that the binding between both proteins involves ion pairing. Furthermore, the formation of an active covalent complex strongly suggests that the lipase-colipase binding site is distinct from the colipase interfacial recognition site as well as from the lipase catalytic site.

【 授权许可】

Unknown