title: UDP-galactopyranose mutase in nematodes.
abstract: Nematodes represent a diverse phylum of both free living and parasitic species. While the species Caenorhabditis elegans is a valuable model organism, parasitic nematodes or helminths pose a serious threat to human health. Indeed, helminths cause many neglected tropical diseases that afflict humans. Nematode glycoconjugates have been implicated in evasive immunomodulation, a hallmark of nematode infections. One monosaccharide residue present in the glycoconjugates of several human pathogens is galactofuranose (Galf). This five-membered ring isomer of galactose has not been detected in mammals, making Galf metabolic enzymes attractive therapeutic targets. The only known pathway for biosynthetic incorporation of Galf into glycoconjugates depends upon generation of the glycosyl donor UDP-Galf by the flavoenzyme uridine 5'-diphosphate (UDP) galactopyranose mutase (UGM or Glf). A putative UGM encoding gene (glf-1) was recently identified in C. elegans. We sought to assess the catalytic activity of the corresponding gene product (CeUGM). CeUGM catalyzes the isomerization of UDP-Galf and UDP-galactopyranose (UDP-Galp). In the presence of enzyme, substrate, and a hydride source, a galactose-N5-FAD adduct was isolated, suggesting the CeUGM flavin adenine dinucleotide (FAD) cofactor serves as a nucleophile in covalent catalysis. Homology modeling and protein variants indicate that CeUGM possesses an active site similar to that of prokaryotic enzymes, despite the low sequence identity (∼15%) between eukaryotic and prokaryotic UGM proteins. Even with the primary sequence differences, heterocyclic UGM inhibitors developed against prokaryotic proteins also inhibit CeUGM activity. We postulate that inhibitors of CeUGM can serve as chemical probes of Galf in nematodes and as anthelmintic leads. The available data suggest that CeUGM facilitates the biosynthetic incorporation of Galf into nematode glycoconjugates through generation of the glycosyl donor UDP-Galf.
authors.affiliation: Department of Biochemistry, University of Wisconsin-Madison, United States.
authors.forename: Darryl A
authors.initials: DA
authors.lastname: Wesener
authors.forename: John F
authors.initials: JF
authors.lastname: May
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: Huffman
authors.forename: Laura L
authors.initials: LL
authors.lastname: Kiessling
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000201:enzymology*; D018360:Crystallography, X-Ray; D006801:Humans; D019751:Intramolecular Transferases / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D019751:Intramolecular Transferases; C098051:UDP-galactopyranose mutase