4EMW

Crystal Structure of Staphylococcus aureus bound with the covalent inhibitor EtVC-CoA

Summary for 4EMW

• Entry DOI: 10.2210/pdb4emw/pdb
• Related: 1YQZ 4EM3 4EM4 4EQR 4EQS 4EQW 4EQX
• Descriptor: Coenzyme A disulfide reductase, FLAVIN-ADENINE DINUCLEOTIDE, MAGNESIUM ION, ... (6 entities in total)
• Functional Keywords: oxidoreductase-oxidoreductase inhibitor complex, oxidoreductase/oxidoreductase inhibitor
• Biological source: Staphylococcus aureus subsp. aureus
• Total number of polymer chains: 2
• Total formula weight: 101505.01

Authors

Edwards, J.S.,Wallace, B.D.,Claiborne, A.,Redinbo, M.R. (deposition date: 2012-04-12, release date: 2012-10-17, Last modification date: 2024-11-20)

Primary citation

Wallace, B.D.,Edwards, J.S.,Wallen, J.R.,Moolman, W.J.,van der Westhuyzen, R.,Strauss, E.,Redinbo, M.R.,Claiborne, A.
Turnover-Dependent Covalent Inactivation of Staphylococcus aureus Coenzyme A-Disulfide Reductase by Coenzyme A-Mimetics: Mechanistic and Structural Insights.
Biochemistry, 51:7699-7711, 2012

PubMed Abstract: Disruption of the unusual thiol-based redox homeostasis mechanisms in Staphylococcus aureus represents a unique opportunity to identify new metabolic processes and new targets for intervention. Targeting uncommon aspects of CoASH biosynthetic and redox functions in S. aureus, the antibiotic CJ-15,801 has recently been demonstrated to be an antimetabolite of the CoASH biosynthetic pathway in this organism; CoAS-mimetics containing α,β-unsaturated sulfone and carboxyl moieties have also been exploited as irreversible inhibitors of S. aureus coenzyme A-disulfide reductase (SaCoADR). In this work we have determined the crystal structures of three of these covalent SaCoADR-inhibitor complexes, prepared by inactivation of wild-type enzyme during turnover. The structures reveal the covalent linkage between the active-site Cys43-S(γ) and C(β) of the vinyl sulfone or carboxyl moiety. The full occupancy of two inhibitor molecules per enzyme dimer, together with kinetic analyses of the wild-type/C43S heterodimer, indicates that half-sites-reactivity is not a factor during normal catalytic turnover. Further, we provide the structures of SaCoADR active-site mutants; in particular, Tyr419'-OH plays dramatic roles in directing intramolecular reduction of the Cys43-SSCoA redox center, in the redox asymmetry observed for the two FAD per dimer in NADPH titrations, and in catalysis. The two conformations observed for the Ser43 side chain in the C43S mutant structure lend support to a conformational switch for Cys43-S(γ) during its catalytic Cys43-SSCoA/Cys43-SH redox cycle. Finally, the structures of the three inhibitor complexes provide a framework for design of more effective inhibitors with therapeutic potential against several major bacterial pathogens.

PubMed: 22954034
DOI: 10.1021/bi301026c

Experimental method

X-RAY DIFFRACTION (2.39 Å)