Enzymatic Synthesis of UDP-Galactofuranose and an Assay for UDP-Galactopyranose Mutase Based on High-Performance Liquid Chromatography

✍ Scribed by Robin Lee; David Monsey; Anthony Weston; Kenneth Duncan; Christopher Rithner; Michael Mcneil

• Publisher: Elsevier Science
• Year: 1996
• Tongue: English
• Weight: 139 KB
• Volume: 242
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.1996.0419

✦ Synopsis

idues has become apparent because they play a crucial A method to prepare UDP-galactofuranose (UDProles in the physiology of significant pathogens. Of spe-Galf ) free of UDP-galactopyranose (UDP-Galp) is decial importance, internal r)5-b-D-Galf and r)6-b-Dscribed. The UDP-Galf is synthesized enzymatically Galf residues are key components of the cell walls of from UDP-Galp using the enzyme UDP-galactopyra-Mycobacterium tuberculosis and other mycobacteria nose mutase. Treatment of UDP-Galp with the enzyme and are essential for the structural integrity of the wall yields an equilibrium mixture of UDP-Galp and UDP-(2). Similarly, an internal b-D-Galf is an integral part Galf in which UDP-Galf is approximately 7%. In spite of the lipid anchor of Leishmania LPG (3) and thus of its low yield, the UDP-Galf is readily purified from necessary for LPG to be expressed on the surface of starting UDP-Galp using a Dionex PA-100 ion exthese organisms. In addition, D-Galf residues are found change HPLC column. The purified UDP-Galf was in lipopeptidophosphoglycans of Trypanosoma cruzi characterized by chemical degradations, by electro-(1), and in glycoinositolphospholipids of Leishmania (4) spray mass spectrometry, and by several nuclear magand Endotrypanum schaundinni (5). They are present netic resonance techniques. In addition, an HPLC in N-linked glycoproteins of Crithidia fasciculata, assay for the enzyme UDP-galactopyranose mutase is Crithidia harmonosa, Leishmania samueli, Herpetopresented that requires 0.5 mg of UDP-Galf per assay monas samuelpessoai, and T. cruzi (1). They are also