title: Structure and Evolution of the Archaeal Lipid Synthesis Enzyme sn-Glycerol-1-phosphate Dehydrogenase.
abstract: One of the most critical events in the origins of cellular life was the development of lipid membranes. Archaea use isoprenoid chains linked via ether bonds to sn-glycerol 1-phosphate (G1P), whereas bacteria and eukaryotes use fatty acids attached via ester bonds to enantiomeric sn-glycerol 3-phosphate. NAD(P)H-dependent G1P dehydrogenase (G1PDH) forms G1P and has been proposed to have played a crucial role in the speciation of the Archaea. We present here, to our knowledge, the first structures of archaeal G1PDH from the hyperthermophilic methanogen Methanocaldococcus jannaschii with bound substrate dihydroxyacetone phosphate, product G1P, NADPH, and Zn(2+) cofactor. We also biochemically characterized the enzyme with respect to pH optimum, cation specificity, and kinetic parameters for dihydroxyacetone phosphate and NAD(P)H. The structures provide key evidence for the reaction mechanism in the stereospecific addition for the NAD(P)H-based pro-R hydrogen transfer and the coordination of the Zn(2+) cofactor during catalysis. Structure-based phylogenetic analyses also provide insight into the origins of G1PDH.
authors.affiliation: From AgResearch Limited, Grasslands Research Centre, Tennent Drive, Private Bag 11008, Palmerston North 4442, New Zealand.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Carbone
authors.affiliation: From AgResearch Limited, Grasslands Research Centre, Tennent Drive, Private Bag 11008, Palmerston North 4442, New Zealand.
authors.forename: Linley R
authors.initials: LR
authors.lastname: Schofield
authors.affiliation: From AgResearch Limited, Grasslands Research Centre, Tennent Drive, Private Bag 11008, Palmerston North 4442, New Zealand.
authors.forename: Yanli
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: From AgResearch Limited, Grasslands Research Centre, Tennent Drive, Private Bag 11008, Palmerston North 4442, New Zealand.
authors.forename: Carrie
authors.initials: C
authors.lastname: Sang
authors.affiliation: From AgResearch Limited, Grasslands Research Centre, Tennent Drive, Private Bag 11008, Palmerston North 4442, New Zealand.
authors.forename: Debjit
authors.initials: D
authors.lastname: Dey
authors.affiliation: From AgResearch Limited, Grasslands Research Centre, Tennent Drive, Private Bag 11008, Palmerston North 4442, New Zealand.
authors.forename: Ingegerd M
authors.initials: IM
authors.lastname: Hannus
authors.affiliation: Institute for Molecular Evolution, Heinrich Heine University, University of Düsseldorf, 40225 Düsseldorf, Germany, and.
authors.forename: William F
authors.initials: WF
authors.lastname: Martin
authors.affiliation: Institute of Fundamental Sciences, Massey University, Palmerston North 4442, New Zealand.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Sutherland-Smith
authors.affiliation: From AgResearch Limited, Grasslands Research Centre, Tennent Drive, Private Bag 11008, Palmerston North 4442, New Zealand, ron.ronimus@agresearch.co.nz.
authors.forename: Ron S
authors.initials: RS
authors.lastname: Ronimus
mesh_terms: D000595:Amino Acid Sequence; D055162:Biocatalysis; D018360:Crystallography, X-Ray; D019143:Evolution, Molecular; D005993:Glycerolphosphate Dehydrogenase / Q000737:chemistry* / Q000235:genetics*; D007700:Kinetics; D008055:Lipids / Q000096:biosynthesis*; D063747:Methanocaldococcus / Q000201:enzymology*; D008969:Molecular Sequence Data; D010802:Phylogeny; D017433:Protein Structure, Secondary; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein; D015032:Zinc / Q000378:metabolism
chemical_list: D008055:Lipids; D005993:Glycerolphosphate Dehydrogenase; C098215:glycerol-1-phosphate dehydrogenase; D015032:Zinc
keywords: archaea; cell membrane; dehydrogenase; evolution; lipid synthesis; phylogenetics; sn-glycerol-1-phosphate dehydrogenase; stereospecific; structural biology; thermophile