Document Detail

Identification of the NAD(P)H binding site of eukaryotic UDP-galactopyranose mutase.
MedLine Citation:
PMID:  23036087     Owner:  NLM     Status:  MEDLINE    
UDP-galactopyranose mutase (UGM) plays an essential role in galactofuranose biosynthesis in microorganisms by catalyzing the conversion of UDP-galactopyranose to UDP-galactofuranose. The enzyme has gained attention recently as a promising target for the design of new antifungal, antitrypanosomal, and antileishmanial agents. Here we report the first crystal structure of UGM complexed with its redox partner NAD(P)H. Kinetic protein crystallography was used to obtain structures of oxidized Aspergillus fumigatus UGM (AfUGM) complexed with NADPH and NADH, as well as reduced AfUGM after dissociation of NADP(+). NAD(P)H binds with the nicotinamide near the FAD isoalloxazine and the ADP moiety extending toward the mobile 200s active site flap. The nicotinamide riboside binding site overlaps that of the substrate galactopyranose moiety, and thus NADPH and substrate binding are mutually exclusive. On the other hand, the pockets for the adenine of NADPH and uracil of the substrate are distinct and separated by only 6 Å, which raises the possibility of designing novel inhibitors that bind both sites. All 12 residues that contact NADP(H) are conserved among eukaryotic UGMs. Residues that form the AMP pocket are absent in bacterial UGMs, which suggests that eukaryotic and bacterial UGMs have different NADP(H) binding sites. The structures address the longstanding question of how UGM binds NAD(P)H and provide new opportunities for drug discovery.
Richa Dhatwalia; Harkewal Singh; Luis M Solano; Michelle Oppenheimer; Reeder M Robinson; Jacob F Ellerbrock; Pablo Sobrado; John J Tanner
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2012-10-19
Journal Detail:
Title:  Journal of the American Chemical Society     Volume:  134     ISSN:  1520-5126     ISO Abbreviation:  J. Am. Chem. Soc.     Publication Date:  2012 Oct 
Date Detail:
Created Date:  2012-10-31     Completed Date:  2013-04-01     Revised Date:  2013-11-06    
Medline Journal Info:
Nlm Unique ID:  7503056     Medline TA:  J Am Chem Soc     Country:  United States    
Other Details:
Languages:  eng     Pagination:  18132-8     Citation Subset:  IM    
Department of Chemistry, University of Missouri-Columbia, Columbia, Missouri 65211, USA.
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MeSH Terms
Aspergillus fumigatus / enzymology*
Binding Sites
Crystallography, X-Ray
Enzyme Activation
Intramolecular Transferases / chemistry*,  genetics,  metabolism
Models, Molecular
Mutagenesis, Site-Directed
NADP / chemistry*,  metabolism
Grant Support
Reg. No./Substance:
53-59-8/NADP; EC 5.4.-/Intramolecular Transferases; EC mutase

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