title: Isolation and characterization of functional Leishmania major virulence factor UDP-galactopyranose mutase.
abstract: Human parasitic pathogens of the genus Leishmania are the causative agents of cutaneous, mucocutaneous, and visceral leishmaniasis. Currently, there are millions of people infected with these diseases and over 50,000 deaths occur annually. Recently, it was shown that the flavin-dependent enzyme UDP-galactopyranose mutase (UGM) is a virulence factor in Leishmania major. UGM catalyzes the conversion of UDP-galactopyranose to UDP-galactofuranose. The product, UDP-galactofuranose, is the only source of galactofuranose which is present on the cell surface of this parasite and has been implicated to be important for host-parasite interactions. The recombinant form of this enzyme was obtained in a soluble and active form. The enzyme was shown to be active only in the reduced state. A k(cat) value of 5 ± 0.2s(-1) and a K(M) value of 87 ± 11 μM were determined with UDP-galactofuranose as the substrate. Different from the dimeric bacterial and tetrameric fungal UGMs, this parasitic enzyme functions as a monomer.
authors.affiliation: Department of Biochemistry, Virginia Tech. Blacksburg, VA 24061, United States.
authors.forename: Michelle
authors.initials: M
authors.lastname: Oppenheimer
authors.forename: Ana L
authors.initials: AL
authors.lastname: Valenciano
authors.forename: Pablo
authors.initials: P
authors.lastname: Sobrado
mesh_terms: D003001:Cloning, Molecular; D006801:Humans; D019751:Intramolecular Transferases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D018320:Leishmania major / Q000201:enzymology* / Q000472:pathogenicity*; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D037521:Virulence Factors / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D011994:Recombinant Proteins; D037521:Virulence Factors; D019751:Intramolecular Transferases; C098051:UDP-galactopyranose mutase