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Article: Refined solution structure of the LpxC-TU-514 complex and pK a analysis of an active site histidine: Insights into the mechanism and inhibitor design

TitleRefined solution structure of the LpxC-TU-514 complex and pK a analysis of an active site histidine: Insights into the mechanism and inhibitor design
Authors
Issue Date2005
PublisherAmerican Chemical Society. The Journal's web site is located at http://pubs.acs.org/biochemistry
Citation
Biochemistry, 2005, v. 44 n. 4, p. 1114-1126 How to Cite?
AbstractLipopolysaccharide, the major constituent of the outer monolayer of the outer membrane of Gram-negative bacteria, is anchored into the membrane through the hydrophobic moiety lipid A, a hexaacylated disaccharide. The zinc-dependent metalloamidase UDP-3-O-acyl-N-acetylglucosamine deacetylase (LpxC) catalyzes the second and committed step in the biosynthesis of lipid A. LpxC shows no homology to mammalian metalloamidases and is essential for cell viability, making it an important target for the development of novel antibacterial compounds. Recent NMR and X-ray studies of the LpxC from Aquifex aeolicus have provided the first structural information about this family of proteins. Insight into the catalytic mechanism and the design of effective inhibitors could be facilitated by more detailed structural and biochemical studies that define substrate-protein interactions and the roles of specific residues in the active site. Here, we report the synthesis of the 13C-labeled substrate-analogue inhibitor TU-514, and the subsequent refinement of the solution structure of the A. aeolicus LpxC-TU-514 complex using residual dipolar couplings. We also reevaluate the catalytic role of an active site histidine, H253, on the basis of both its pK a as determined by NMR titration and pH-dependent kinetic analyses. These results provide a structural basis for the design of more potent LpxC inhibitors than those that are currently available.
Persistent Identifierhttp://hdl.handle.net/10722/167891
ISSN
2015 Impact Factor: 2.876
2015 SCImago Journal Rankings: 1.769
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorCoggins, BEen_US
dc.contributor.authorMcclerren, ALen_US
dc.contributor.authorJiang, Len_US
dc.contributor.authorLi, Xen_US
dc.contributor.authorRudolph, Jen_US
dc.contributor.authorHindsgaul, Oen_US
dc.contributor.authorRaetz, CRHen_US
dc.contributor.authorZhou, Pen_US
dc.date.accessioned2012-10-08T03:12:36Z-
dc.date.available2012-10-08T03:12:36Z-
dc.date.issued2005en_US
dc.identifier.citationBiochemistry, 2005, v. 44 n. 4, p. 1114-1126en_US
dc.identifier.issn0006-2960en_US
dc.identifier.urihttp://hdl.handle.net/10722/167891-
dc.description.abstractLipopolysaccharide, the major constituent of the outer monolayer of the outer membrane of Gram-negative bacteria, is anchored into the membrane through the hydrophobic moiety lipid A, a hexaacylated disaccharide. The zinc-dependent metalloamidase UDP-3-O-acyl-N-acetylglucosamine deacetylase (LpxC) catalyzes the second and committed step in the biosynthesis of lipid A. LpxC shows no homology to mammalian metalloamidases and is essential for cell viability, making it an important target for the development of novel antibacterial compounds. Recent NMR and X-ray studies of the LpxC from Aquifex aeolicus have provided the first structural information about this family of proteins. Insight into the catalytic mechanism and the design of effective inhibitors could be facilitated by more detailed structural and biochemical studies that define substrate-protein interactions and the roles of specific residues in the active site. Here, we report the synthesis of the 13C-labeled substrate-analogue inhibitor TU-514, and the subsequent refinement of the solution structure of the A. aeolicus LpxC-TU-514 complex using residual dipolar couplings. We also reevaluate the catalytic role of an active site histidine, H253, on the basis of both its pK a as determined by NMR titration and pH-dependent kinetic analyses. These results provide a structural basis for the design of more potent LpxC inhibitors than those that are currently available.en_US
dc.languageengen_US
dc.publisherAmerican Chemical Society. The Journal's web site is located at http://pubs.acs.org/biochemistryen_US
dc.relation.ispartofBiochemistryen_US
dc.subject.meshAmidohydrolases - Antagonists & Inhibitors - Chemistry - Geneticsen_US
dc.subject.meshAquifoliaceae - Enzymology - Geneticsen_US
dc.subject.meshBinding Sites - Geneticsen_US
dc.subject.meshCatalysisen_US
dc.subject.meshConserved Sequence - Geneticsen_US
dc.subject.meshCrystallography, X-Rayen_US
dc.subject.meshEnzyme Inhibitors - Chemical Synthesisen_US
dc.subject.meshGlycolipids - Chemical Synthesisen_US
dc.subject.meshHexoses - Chemistryen_US
dc.subject.meshHistidine - Chemistry - Geneticsen_US
dc.subject.meshHydrogen-Ion Concentrationen_US
dc.subject.meshMutagenesis, Site-Directeden_US
dc.subject.meshNuclear Magnetic Resonance, Biomolecular - Methodsen_US
dc.subject.meshProtonsen_US
dc.subject.meshSolutionsen_US
dc.subject.meshSubstrate Specificity - Geneticsen_US
dc.subject.meshThermodynamicsen_US
dc.titleRefined solution structure of the LpxC-TU-514 complex and pK a analysis of an active site histidine: Insights into the mechanism and inhibitor designen_US
dc.typeArticleen_US
dc.identifier.emailLi, X:xuechenl@hku.hken_US
dc.identifier.authorityLi, X=rp00742en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1021/bi047820zen_US
dc.identifier.pmid15667205-
dc.identifier.scopuseid_2-s2.0-13444287735en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-13444287735&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume44en_US
dc.identifier.issue4en_US
dc.identifier.spage1114en_US
dc.identifier.epage1126en_US
dc.identifier.isiWOS:000226594800004-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridCoggins, BE=6507075734en_US
dc.identifier.scopusauthoridMcClerren, AL=7801327115en_US
dc.identifier.scopusauthoridJiang, L=36675320800en_US
dc.identifier.scopusauthoridLi, X=24168958800en_US
dc.identifier.scopusauthoridRudolph, J=7103011865en_US
dc.identifier.scopusauthoridHindsgaul, O=7102966220en_US
dc.identifier.scopusauthoridRaetz, CRH=7102514726en_US
dc.identifier.scopusauthoridZhou, P=7401848622en_US

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