title: Discrimination between closed and open forms of lipases using electrophoretic techniques.
abstract: The enhanced catalytic activity of lipases is often associated with structural changes. The three-dimensional (3D) structures showed that the covalently inhibited lipases exist under their open conformations, in contrast to their native closed forms. We studied the inhibition of various lipases--human and dog gastric lipases, human pancreatic lipase, and Humicola lanuginosa lipase--by the octyl-undecyl phosphonate inhibitor, and we measured the subsequent modifications of their respective electrophoretic mobility. Furthermore, the experimental values of the isoelectric points found for the native (closed) and inhibited (open) lipases are in agreement with theoretical calculations based on the electrostatic potential. We concluded that there is a significant difference in the isoelectric points between the closed (native) and open (inhibited) conformations of the four lipases investigated. Thus, analysis of the electrophoretic pattern is proposed as an easy experimental tool to differentiate between a closed and an open form of a given lipase.
authors.affiliation: UPR 9025 du CNRS, Laboratoire de Lipolyse Enzymatique, 31 Chemin Joseph-Aiguier, 13402 Marseille Cedex 20, France.
authors.forename: N
authors.initials: N
authors.lastname: Miled
authors.forename: M
authors.initials: M
authors.lastname: Riviere
authors.forename: J F
authors.initials: JF
authors.lastname: Cavalier
authors.forename: G
authors.initials: G
authors.lastname: Buono
authors.forename: L
authors.initials: L
authors.lastname: Berti
authors.forename: R
authors.initials: R
authors.lastname: Verger
mesh_terms: D000818:Animals; D001203:Ascomycota / Q000201:enzymology; D004285:Dogs; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005960:Glucosides / Q000494:pharmacology; D006801:Humans; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D008049:Lipase / Q000037:antagonists & inhibitors / Q000737:chemistry*; D010179:Pancreas / Q000201:enzymology
chemical_list: D004791:Enzyme Inhibitors; D005960:Glucosides; D008049:Lipase; C018619:octyl-beta-D-glucoside; C485982:LIPF protein, human