title: Leishmania UDP-sugar pyrophosphorylase: the missing link in galactose salvage?
abstract: The Leishmania parasite glycocalyx is rich in galactose-containing glycoconjugates that are synthesized by specific glycosyltransferases that use UDP-galactose as a glycosyl donor. UDP-galactose biosynthesis is thought to be predominantly a de novo process involving epimerization of the abundant nucleotide sugar UDP-glucose by the UDP-glucose 4-epimerase, although galactose salvage from the environment has been demonstrated for Leishmania major. Here, we present the characterization of an L. major UDP-sugar pyrophosphorylase able to reversibly activate galactose 1-phosphate into UDP-galactose thus proving the existence of the Isselbacher salvage pathway in this parasite. The ordered bisubstrate mechanism and high affinity of the enzyme for UTP seem to favor the synthesis of nucleotide sugar rather than their pyrophosphorolysis. Although L. major UDP-sugar pyrophosphorylase preferentially activates galactose 1-phosphate and glucose 1-phosphate, the enzyme is able to act on a variety of hexose 1-phosphates as well as pentose 1-phosphates but not hexosamine 1-phosphates and hence presents a broad in vitro specificity. The newly identified enzyme exhibits a low but significant homology with UDP-glucose pyrophosphorylases and conserved in particular is the pyrophosphorylase consensus sequence and residues involved in nucleotide and phosphate binding. Saturation transfer difference NMR spectroscopy experiments confirm the importance of these moieties for substrate binding. The described leishmanial enzyme is closely related to plant UDP-sugar pyrophosphorylases and presents a similar substrate specificity suggesting their common origin.
authors.affiliation: Department of Cellular Chemistry, Hannover Medical School, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Damerow
authors.forename: Anne-Christin
authors.initials: AC
authors.lastname: Lamerz
authors.forename: Thomas
authors.initials: T
authors.lastname: Haselhorst
authors.forename: Jana
authors.initials: J
authors.lastname: Führing
authors.forename: Patricia
authors.initials: P
authors.lastname: Zarnovican
authors.forename: Mark
authors.initials: M
authors.lastname: von Itzstein
authors.forename: Françoise H
authors.initials: FH
authors.lastname: Routier
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019143:Evolution, Molecular; D005695:Galactosephosphates / Q000235:genetics / Q000378:metabolism*; D005958:Glucosephosphates / Q000235:genetics / Q000378:metabolism; D019276:Glycocalyx / Q000201:enzymology / Q000235:genetics; D018320:Leishmania major / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D009713:Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism*; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity / Q000502:physiology; D014530:Uridine Diphosphate / Q000235:genetics / Q000378:metabolism*; D014544:Uridine Triphosphate / Q000235:genetics / Q000378:metabolism
chemical_list: D005695:Galactosephosphates; D005958:Glucosephosphates; D015800:Protozoan Proteins; C029973:galactose-1-phosphate; D014530:Uridine Diphosphate; C031590:glucose-1-phosphate; D009713:Nucleotidyltransferases; D014544:Uridine Triphosphate