title: Influence of the conformational flexibility on the kinetics and dimerisation process of two Candida rugosa lipase isoenzymes.
abstract: We have investigated the interfacial activation process of two isoenzymes from Candida rugosa (Lip1 and Lip3) using triacetin as substrate. Kinetics were coupled to inhibition experiments in order to analyse the transition between the open and closed conformers. This process was slow, particularly for Lip1, in the absence of an interface provided by the substrate or a detergent. Dimers of Lip3 were also purified and their catalytic action was closer to that of a typical esterase. In spite of the high sequence homology between Lip1 and Lip3, small changes enhance hydrophobicity in the binding pocket of Lip3 and increase the flexibility of its flap. We postulated that these factors account for the higher tendency of Lip3 to dimerise fixing its open conformation.
authors.affiliation: Area de Bioquímica y Biologíca Molecular, facultade de Ciencias de Ourense, Universidade de Vigo, Spain.
authors.forename: M A
authors.initials: MA
authors.lastname: Pernas
authors.forename: C
authors.initials: C
authors.lastname: López
authors.forename: M L
authors.initials: ML
authors.lastname: Rúa
authors.forename: J
authors.initials: J
authors.lastname: Hermoso
mesh_terms: D001665:Binding Sites; D002175:Candida / Q000201:enzymology*; D002850:Chromatography, Gel; D019281:Dimerization; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008049:Lipase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D008958:Models, Molecular; D008970:Molecular Weight; D010261:Paraoxon / Q000494:pharmacology; D018583:Pliability; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D014215:Triacetin / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D007527:Isoenzymes; D008049:Lipase; D010261:Paraoxon; D014215:Triacetin