title: Structural basis for catalysis in a CDP-alcohol phosphotransferase.
abstract: The CDP-alcohol phosphotransferase (CDP-AP) family of integral membrane enzymes catalyses the transfer of a substituted phosphate group from a CDP-linked donor to an alcohol acceptor. This is an essential reaction for phospholipid biosynthesis across all kingdoms of life, and it is catalysed solely by CDP-APs. Here we report the 2.0 Å resolution crystal structure of a representative CDP-AP from Archaeoglobus fulgidus. The enzyme (AF2299) is a homodimer, with each protomer consisting of six transmembrane helices and an N-terminal cytosolic domain. A polar cavity within the membrane accommodates the active site, lined with the residues from an absolutely conserved CDP-AP signature motif (D(1)xxD(2)G(1)xxAR...G(2)xxxD(3)xxxD(4)). Structures in the apo, CMP-bound, CDP-bound and CDP-glycerol-bound states define functional roles for each of these eight conserved residues and allow us to propose a sequential, base-catalysed mechanism universal for CDP-APs, in which the fourth aspartate (D4) acts as the catalytic base.
authors.affiliation: 1] Department of Physiology and Cellular Biophysics, Columbia University, New York, New York 10032, USA [2].
authors.forename: Giuliano
authors.initials: G
authors.lastname: Sciara
authors.affiliation: 1] Department of Biochemistry and Molecular Biophysics, Columbia University, New York, New York 10032, USA [2].
authors.forename: Oliver B
authors.initials: OB
authors.lastname: Clarke
authors.affiliation: 1] Department of Physiology and Cellular Biophysics, Columbia University, New York, New York 10032, USA [2].
authors.forename: David
authors.initials: D
authors.lastname: Tomasek
authors.affiliation: New York Consortium on Membrane Protein Structure, New York Structural Biology Center, 89 Convent Avenue, New York, New York 10027, USA.
authors.forename: Brian
authors.initials: B
authors.lastname: Kloss
authors.affiliation: New York Consortium on Membrane Protein Structure, New York Structural Biology Center, 89 Convent Avenue, New York, New York 10027, USA.
authors.forename: Shantelle
authors.initials: S
authors.lastname: Tabuso
authors.affiliation: Department of Physiology and Cellular Biophysics, Columbia University, New York, New York 10032, USA.
authors.forename: Rushelle
authors.initials: R
authors.lastname: Byfield
authors.affiliation: Department of Physiology and Cellular Biophysics, Columbia University, New York, New York 10032, USA.
authors.forename: Raphael
authors.initials: R
authors.lastname: Cohn
authors.affiliation: Department of Chemistry and Chemical Biology, Cornell University, NE-CAT, Advanced Photon Source, Argonne, Illinois 60439, USA.
authors.forename: Surajit
authors.initials: S
authors.lastname: Banerjee
authors.affiliation: Department of Chemistry and Chemical Biology, Cornell University, NE-CAT, Advanced Photon Source, Argonne, Illinois 60439, USA.
authors.forename: Kanagalaghatta R
authors.initials: KR
authors.lastname: Rajashankar
authors.affiliation: Department of Environmental Health Sciences, Mailman School of Public Health, Columbia University, New York, New York 10032, USA.
authors.forename: Vesna
authors.initials: V
authors.lastname: Slavkovic
authors.affiliation: Department of Environmental Health Sciences, Mailman School of Public Health, Columbia University, New York, New York 10032, USA.
authors.forename: Joseph H
authors.initials: JH
authors.lastname: Graziano
authors.affiliation: Department of Biochemistry and Molecular Biophysics, Columbia University, New York, New York 10032, USA.
authors.forename: Lawrence
authors.initials: L
authors.lastname: Shapiro
authors.affiliation: Department of Physiology and Cellular Biophysics, Columbia University, New York, New York 10032, USA.
authors.forename: Filippo
authors.initials: F
authors.lastname: Mancia
mesh_terms: D000438:Alcohols / Q000378:metabolism*; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019611:Archaeoglobus fulgidus / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D001665:Binding Sites; D055162:Biocatalysis; D020134:Catalytic Domain; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment
chemical_list: D000438:Alcohols; D019843:Archaeal Proteins; D017853:Phosphotransferases (Alcohol Group Acceptor)