A cross‐linked complex between horse pancreatic lipase and colipase
- 6 November 1989
- journal article
- Published by Wiley in FEBS Letters
- Vol. 257 (2), 443-446
- https://doi.org/10.1016/0014-5793(89)81592-3
Abstract
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.Keywords
This publication has 18 references indexed in Scilit:
- Crystallization and preliminary X-ray study of horse pancreatic lipaseJournal of Molecular Biology, 1989
- Purification of pancreatic carboxylic-ester hydrolase by immunoaffinity and its application to the human bile-salt-stimulated lipaseBiochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism, 1988
- Spectrofluorimetric study of the bile salt micelle binding site of pig and horse colipasesBiochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1986
- “Western Blotting”: Electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein AAnalytical Biochemistry, 1981
- Small-angle neutron scattering study of the association between porcine pancreatic colipase and taurodeoxycholate micellesJournal of Molecular Biology, 1980
- Chemical modifications of pancreatic colipaseBiochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism, 1977
- Role of colipase in the interfacial adsorption of pancreatic lipase at hydrophilic interfacesFEBS Letters, 1975
- Interactions of Colipase with Bile Salt MicellesEuropean Journal of Biochemistry, 1975
- On porcine pancreatic colipase: Large scale purification and some propertiesBiochemical and Biophysical Research Communications, 1973
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970