title: Importance of the lid and cap domains for the catalytic activity of gastric lipases.
abstract: Human gastric lipase (HGL) is an enzyme secreted by the stomach, which is stable and active despite the highly acidic environment. It has been clearly established that this enzyme is responsible for 30% of the fat digestion processes occurring in human. This globular protein belongs to the alpha/beta hydrolase fold family and its catalytic serine is deeply buried under a domain called the extrusion domain, which is composed of a 'cap' domain and a segment consisting of 58 residues, which can be defined as a lid. The exact roles played by the cap and the lid domains during the catalytic step have not yet been elucidated. We have recently solved the crystal structure of the open form of the dog gastric lipase in complex with a covalent inhibitor. The detergent molecule and the inhibitor were mimicking a triglyceride substrate that would interact with residues belonging to both the cap and the lid domains. In this study, we have investigated the role of the cap and the lid domains, using site-directed mutagenesis procedures. We have produced truncated mutants lacking the lid and the cap. After expressing these mutants and purifying them, their activity was found to have decreased drastically in comparison with the wild type HGL. The lid and the cap domains play an important role in the catalytic reaction mechanism. Based on these results and the structural data (open form of DGL), we have pointed out the cap and the lid residues involved in the binding with the lipidic substrate.
authors.affiliation: Laboratoire de Lipolyse Enzymatique, UPR 9025 de l'IFR-1 du CNRS, 31 Chemin Joseph Aiguier, Marseille 20 13402, France.
authors.forename: N
authors.initials: N
authors.lastname: Miled
authors.forename: C
authors.initials: C
authors.lastname: Bussetta
authors.forename: A
authors.initials: A
authors.lastname: De caro
authors.forename: M
authors.initials: M
authors.lastname: Rivière
authors.forename: L
authors.initials: L
authors.lastname: Berti
authors.forename: S
authors.initials: S
authors.lastname: Canaan
mesh_terms: D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D002384:Catalysis; D018604:Epitope Mapping; D006801:Humans; D007700:Kinetics; D008049:Lipase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D013270:Stomach / Q000201:enzymology*
chemical_list: D000911:Antibodies, Monoclonal; D008049:Lipase; C485982:LIPF protein, human