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A cross‐linked complex between horse pancreatic lipase and colipase
Author(s) -
Chaillan Catherine,
Rogalska Ewa,
Chapus Catherine,
Lombardo Dominique
Publication year - 1989
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(89)81592-3
Subject(s) - colipase , lipase , carbodiimide , chemistry , covalent bond , biochemistry , triacylglycerol lipase , enzyme , organic chemistry
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.