title: Structural basis for selective recognition of acyl chains by the membrane-associated acyltransferase PatA.
abstract: The biosynthesis of phospholipids and glycolipids are critical pathways for virtually all cell membranes. PatA is an essential membrane associated acyltransferase involved in the biosynthesis of mycobacterial phosphatidyl-myo-inositol mannosides (PIMs). The enzyme transfers a palmitoyl moiety from palmitoyl-CoA to the 6-position of the mannose ring linked to 2-position of inositol in PIM1/PIM2. We report here the crystal structures of PatA from Mycobacterium smegmatis in the presence of its naturally occurring acyl donor palmitate and a nonhydrolyzable palmitoyl-CoA analog. The structures reveal an α/β architecture, with the acyl chain deeply buried into a hydrophobic pocket that runs perpendicular to a long groove where the active site is located. Enzyme catalysis is mediated by an unprecedented charge relay system, which markedly diverges from the canonical HX4D motif. Our studies establish the mechanistic basis of substrate/membrane recognition and catalysis for an important family of acyltransferases, providing exciting possibilities for inhibitor design.
authors.affiliation: Unidad de Biofísica, Consejo Superior de Investigaciones Científicas-Universidad del País Vasco/Euskal Herriko Unibertsitatea (CSIC,UPV/EHU), Barrio Sarriena s/n, Leioa, 48940 Bizkaia, Spain.|Departamento de Bioquímica, Universidad del País Vasco, Leioa, 48940 Bizkaia, Spain.|IKERBASQUE, Basque Foundation for Science, 48013 Bilbao, Spain.
authors.forename: David
authors.identifier: http://orcid.org/0000-0003-2904-8203
authors.initials: D
authors.lastname: Albesa-Jové
authors.affiliation: Department of Biochemistry, Faculty of Natural Sciences, Comenius University in Bratislava, 84215 Bratislava, Slovakia.
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Svetlíková
authors.affiliation: Unidad de Biofísica, Consejo Superior de Investigaciones Científicas-Universidad del País Vasco/Euskal Herriko Unibertsitatea (CSIC,UPV/EHU), Barrio Sarriena s/n, Leioa, 48940 Bizkaia, Spain.|Departamento de Bioquímica, Universidad del País Vasco, Leioa, 48940 Bizkaia, Spain.
authors.forename: Montse
authors.initials: M
authors.lastname: Tersa
authors.affiliation: Unidad de Biofísica, Consejo Superior de Investigaciones Científicas-Universidad del País Vasco/Euskal Herriko Unibertsitatea (CSIC,UPV/EHU), Barrio Sarriena s/n, Leioa, 48940 Bizkaia, Spain.|Departamento de Bioquímica, Universidad del País Vasco, Leioa, 48940 Bizkaia, Spain.
authors.forename: Enea
authors.initials: E
authors.lastname: Sancho-Vaello
authors.affiliation: Unidad de Biofísica, Consejo Superior de Investigaciones Científicas-Universidad del País Vasco/Euskal Herriko Unibertsitatea (CSIC,UPV/EHU), Barrio Sarriena s/n, Leioa, 48940 Bizkaia, Spain.|Departamento de Bioquímica, Universidad del País Vasco, Leioa, 48940 Bizkaia, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Carreras-González
authors.affiliation: Université d'Orléans et CNRS, ICOA, UMR 7311, F-45067 Orléans, France.
authors.forename: Pascal
authors.initials: P
authors.lastname: Bonnet
authors.affiliation: Unidad de Biofísica, Consejo Superior de Investigaciones Científicas-Universidad del País Vasco/Euskal Herriko Unibertsitatea (CSIC,UPV/EHU), Barrio Sarriena s/n, Leioa, 48940 Bizkaia, Spain.|Departamento de Bioquímica, Universidad del País Vasco, Leioa, 48940 Bizkaia, Spain.
authors.forename: Pedro
authors.initials: P
authors.lastname: Arrasate
authors.affiliation: Unidad de Biofísica, Consejo Superior de Investigaciones Científicas-Universidad del País Vasco/Euskal Herriko Unibertsitatea (CSIC,UPV/EHU), Barrio Sarriena s/n, Leioa, 48940 Bizkaia, Spain.|Departamento de Bioquímica, Universidad del País Vasco, Leioa, 48940 Bizkaia, Spain.
authors.forename: Ander
authors.initials: A
authors.lastname: Eguskiza
authors.affiliation: Mycobacteria Research Laboratories, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado 80523-1682, USA.
authors.forename: Shiva K
authors.initials: SK
authors.lastname: Angala
authors.affiliation: Unidad de Biofísica, Consejo Superior de Investigaciones Científicas-Universidad del País Vasco/Euskal Herriko Unibertsitatea (CSIC,UPV/EHU), Barrio Sarriena s/n, Leioa, 48940 Bizkaia, Spain.|Departamento de Bioquímica, Universidad del País Vasco, Leioa, 48940 Bizkaia, Spain.
authors.forename: Javier O
authors.initials: JO
authors.lastname: Cifuente
authors.affiliation: Department of Biochemistry, Faculty of Natural Sciences, Comenius University in Bratislava, 84215 Bratislava, Slovakia.
authors.forename: Jana
authors.initials: J
authors.lastname: Korduláková
authors.affiliation: Mycobacteria Research Laboratories, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado 80523-1682, USA.
authors.forename: Mary
authors.initials: M
authors.lastname: Jackson
authors.affiliation: Department of Biochemistry, Faculty of Natural Sciences, Comenius University in Bratislava, 84215 Bratislava, Slovakia.
authors.forename: Katarína
authors.initials: K
authors.lastname: Mikušová
authors.affiliation: Unidad de Biofísica, Consejo Superior de Investigaciones Científicas-Universidad del País Vasco/Euskal Herriko Unibertsitatea (CSIC,UPV/EHU), Barrio Sarriena s/n, Leioa, 48940 Bizkaia, Spain.|Departamento de Bioquímica, Universidad del País Vasco, Leioa, 48940 Bizkaia, Spain.|IKERBASQUE, Basque Foundation for Science, 48013 Bilbao, Spain.|Structural Biology Unit, CIC bioGUNE, Bizkaia Technology Park, 48160 Derio, Spain.
authors.forename: Marcelo E
authors.initials: ME
authors.lastname: Guerin
mesh_terms: D000217:Acyltransferases / Q000737:chemistry / Q000378:metabolism*; D002384:Catalysis; D020134:Catalytic Domain; D002462:Cell Membrane / Q000378:metabolism; D018360:Crystallography, X-Ray; D008362:Mannosides / Q000096:biosynthesis; D020102:Mycobacterium smegmatis / Q000737:chemistry / Q000378:metabolism*; D010168:Palmitates / Q000378:metabolism; D010171:Palmitoyl Coenzyme A / Q000378:metabolism; D010716:Phosphatidylinositols / Q000096:biosynthesis; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary
chemical_list: D008362:Mannosides; D010168:Palmitates; D010716:Phosphatidylinositols; D010171:Palmitoyl Coenzyme A; D000217:Acyltransferases