title: Structural and functional analysis of the gpsA gene product of Archaeoglobus fulgidus: a glycerol-3-phosphate dehydrogenase with an unusual NADP+ preference.
abstract: NAD(+)-dependent glycerol-3-phosphate dehydrogenase (G3PDH) is generally absent in archaea, because archaea, unlike eukaryotes and eubacteria, utilize glycerol-1-phosphate instead of glycerol-3-phosphate for the biosynthesis of membrane lipids. Surprisingly, the genome of the hyperthermophilic archaeon Archaeoglobus fulgidus comprises a G3PDH ortholog, gpsA, most likely due to horizontal gene transfer from a eubacterial organism. Biochemical characterization proved G3PDH-like activity of the recombinant gpsA gene product. However, unlike other G3PDHs, the up to 85 degrees C thermostable A. fulgidus G3PDH exerted a 15-fold preference for NADPH over NADH. The A. fulgidus G3PDH bears the hallmarks of adaptation to halotolerance and thermophilicity, because its 1.7-A crystal structure showed a high surface density for negative charges and 10 additional intramolecular salt bridges compared to a mesophilic G3PDH structure. Whereas all amino acid residues required for dihydroxyacetone phosphate binding and reductive catalysis are highly conserved, the binding site for the adenine moiety of the NAD(P) cosubstrate shows a structural variation that reflects the observed NADPH preference, for example, by a putative salt bridge between R49 and the 2'-phosphate.
authors.affiliation: Max-Planck-Institut für terrestrische Mikrobiologie and Laboratorium für Mikrobiologie, Fachbereich Biologie, Philipps Universität, Marburg, Germany.
authors.forename: Shin-Ichi
authors.initials: S
authors.lastname: Sakasegawa
authors.forename: Christoph H
authors.initials: CH
authors.lastname: Hagemeier
authors.forename: Rudolf K
authors.initials: RK
authors.lastname: Thauer
authors.forename: Lars-O
authors.initials: LO
authors.lastname: Essen
authors.forename: Seigo
authors.initials: S
authors.lastname: Shima
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019611:Archaeoglobus fulgidus / Q000201:enzymology* / Q000235:genetics; D001665:Binding Sites; D018360:Crystallography, X-Ray; D005993:Glycerolphosphate Dehydrogenase / Q000737:chemistry* / Q000502:physiology*; D007894:Leishmania mexicana / Q000201:enzymology; D008969:Molecular Sequence Data; D009249:NADP / Q000378:metabolism*; D011487:Protein Conformation; D016415:Sequence Alignment
chemical_list: D009249:NADP; D005993:Glycerolphosphate Dehydrogenase; C096203:glycerol-3-phosphate dehydrogenase (NAD(P)+)