title: Crystal structure of the Saccharomyces cerevisiae monoglyceride lipase Yju3p.
abstract: Monoglyceride lipases (MGLs) are a group of α/β-hydrolases that catalyze the hydrolysis of monoglycerides (MGs) into free fatty acids and glycerol. This reaction serves different physiological functions, namely in the last step of phospholipid and triglyceride degradation, in mammalian endocannabinoid and arachidonic acid metabolism, and in detoxification processes in microbes. Previous crystal structures of MGLs from humans and bacteria revealed conformational plasticity in the cap region of this protein and gave insight into substrate binding. In this study, we present the structure of a MGL from Saccharomyces cerevisiae called Yju3p in its free form and in complex with a covalently bound substrate analog mimicking the tetrahedral intermediate of MG hydrolysis. These structures reveal a high conservation of the overall shape of the MGL cap region and also provide evidence for conformational changes in the cap of Yju3p. The complex structure reveals that, despite the high structural similarity, Yju3p seems to have an additional opening to the substrate binding pocket at a different position compared to human and bacterial MGL. Substrate specificities towards MGs with saturated and unsaturated alkyl chains of different lengths were tested and revealed highest activity towards MG containing a C18:1 fatty acid.
authors: Institute of Molecular Biosciences, University of Graz, Humboldtstraße 50/3, 8010 Graz, Austria.
authors: Philipp
authors: P
authors: Aschauer
authors: Institute of Molecular Biosciences, University of Graz, Humboldtstraße 50/3, 8010 Graz, Austria.
authors: Srinivasan
authors: S
authors: Rengachari
authors: Institute of Molecular Biosciences, University of Graz, Humboldtstraße 50/3, 8010 Graz, Austria.
authors: Joerg
authors: J
authors: Lichtenegger
authors: Research Unit Functional Proteomics and Metabolic Pathways, Institute of Pathology, Medical University of Graz, Stiftingtalstrasse 24, 8010 Graz, Austria; Omics Center Graz, BioTechMed-Graz, Stiftingtalstrasse 24, 8010 Graz, Austria.
authors: Matthias
authors: M
authors: Schittmayer
authors: Institute of Molecular Biosciences, University of Graz, Humboldtstraße 50/3, 8010 Graz, Austria.
authors: Krishna Mohan Padmanabha
authors: KM
authors: Das
authors: Institute of Organic Chemistry, Graz University of Technology, Stremayrgasse 9, 8010 Graz, Austria.
authors: Nicole
authors: N
authors: Mayer
authors: Institute of Organic Chemistry, Graz University of Technology, Stremayrgasse 9, 8010 Graz, Austria.
authors: Rolf
authors: R
authors: Breinbauer
authors: Research Unit Functional Proteomics and Metabolic Pathways, Institute of Pathology, Medical University of Graz, Stiftingtalstrasse 24, 8010 Graz, Austria; Omics Center Graz, BioTechMed-Graz, Stiftingtalstrasse 24, 8010 Graz, Austria.
authors: Ruth
authors: R
authors: Birner-Gruenberger
authors: Institute of Molecular Biosciences, University of Graz, Humboldtstraße 50/3, 8010 Graz, Austria; ACIB - Austrian Centre of Industrial Biotechnology, Petersgasse 14, 8010 Graz, Austria.
authors: Christian C
authors: CC
authors: Gruber
authors: Institute of Molecular Biosciences, University of Graz, Humboldtstraße 50/3, 8010 Graz, Austria.
authors: Robert
authors: R
authors: Zimmermann
authors: Institute of Molecular Biosciences, University of Graz, Humboldtstraße 50/3, 8010 Graz, Austria.
authors: Karl
authors: K
authors: Gruber
authors: Institute of Molecular Biosciences, University of Graz, Humboldtstraße 50/3, 8010 Graz, Austria. Electronic address: m.oberer@uni-graz.at.
authors: Monika
authors: M
authors: Oberer
mesh_terms: D001665:Binding Sites; D002384:Catalysis; D003001:Cloning, Molecular; D003460:Crystallization; D006868:Hydrolysis; D056004:Molecular Dynamics Simulation; D008994:Monoacylglycerol Lipases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D050178:Monoglycerides / Q000737:chemistry* / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: D050178:Monoglycerides; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D008994:Monoacylglycerol Lipases; C552818:Yju3 protein, S cerevisiae
keywords: Crystal structure; Lipase cap; Lipase–substrate-complex; Monoacylglycerol lipase; Monoglyceride lipase; Substrate specificity