6CLU

Staphylococcus aureus Dihydropteroate Synthase (saDHPS) F17L E208K double mutant structure

Summary for 6CLU

• Entry DOI: 10.2210/pdb6clu/pdb
• Descriptor: Dihydropteroate synthase (2 entities in total)
• Functional Keywords: antibiotic resistance mutations, sulfonamide resistance, antimicrobial protein
• Biological source: Staphylococcus aureus
• Total number of polymer chains: 4
• Total formula weight: 129627.74

Authors

Gajewski, S.,Griffith, E.C.,Wu, Y.,White, S.W. (deposition date: 2018-03-02, release date: 2018-08-22, Last modification date: 2024-03-13)

Primary citation

Griffith, E.C.,Wallace, M.J.,Wu, Y.,Kumar, G.,Gajewski, S.,Jackson, P.,Phelps, G.A.,Zheng, Z.,Rock, C.O.,Lee, R.E.,White, S.W.
The Structural and Functional Basis for Recurring Sulfa Drug Resistance Mutations inStaphylococcus aureusDihydropteroate Synthase.
Front Microbiol, 9:1369-1369, 2018

PubMed Abstract: Staphylococcal species are a leading cause of bacterial drug-resistant infections and associated mortality. One strategy to combat bacterial drug resistance is to revisit compromised targets, and to circumvent resistance mechanisms using structure-assisted drug discovery. The folate pathway is an ideal candidate for this approach. Antifolates target an essential metabolic pathway, and the necessary detailed structural information is now available for most enzymes in this pathway. Dihydropteroate synthase (DHPS) is the target of the sulfonamide class of drugs, and its well characterized mechanism facilitates detailed analyses of how drug resistance has evolved. Here, we surveyed clinical genetic sequencing data in to distinguish natural amino acid variations in DHPS from those that are associated with sulfonamide resistance. Five mutations were identified, F17L, S18L, T51M, E208K, and KE257_dup. Their contribution to resistance and their cost to the catalytic properties of DHPS were evaluated using a combination of biochemical, biophysical and microbiological susceptibility studies. These studies show that F17L, S18L, and T51M directly lead to sulfonamide resistance while unexpectedly increasing susceptibility to trimethoprim, which targets the downstream enzyme dihydrofolate reductase. The secondary mutations E208K and KE257_dup restore trimethoprim susceptibility closer to wild-type levels while further increasing sulfonamide resistance. Structural studies reveal that these mutations appear to selectively disfavor the binding of the sulfonamides by sterically blocking an outer ring moiety that is not present in the substrate. This emphasizes that new inhibitors must be designed that strictly stay within the substrate volume in the context of the transition state.

PubMed: 30065703
DOI: 10.3389/fmicb.2018.01369

Experimental method

X-RAY DIFFRACTION (1.95 Å)