PDBsum entry 5isp

Oxidoreductase

PDB id: 5isp

Contents

Protein chain

157 a.a.

Ligands

NAP

U06

GOL

Waters ×100

PDB id:

5isp

Name:

Oxidoreductase

Title:

Staphylococcus aureus f98y dihydrofolate reductase mutant complexed with beta-NADPH and 3'-(3-(2,4-diamino-6-ethylpyrimidin-5-yl)prop-2- yn-1-yl)-4'-methoxy-[1,1'-biphenyl]-4-carboxylic acid (ucp1106)

Structure:

Dihydrofolate reductase. Chain: x. Synonym: dhfr. Engineered: yes. Mutation: yes

Source:

Staphylococcus aureus. Organism_taxid: 1280. Atcc: 43300. Gene: fola. Expressed in: escherichia coli. Expression_system_taxid: 469008.

Resolution:

1.84Å R-factor: 0.182 R-free: 0.210

Authors:

A.C.Anderson,S.M.Reeve

Key ref:

S.M.Reeve et al. (2016). Charged Propargyl-Linked Antifolates Reveal Mechanisms of Antifolate Resistance and Inhibit Trimethoprim-Resistant MRSA Strains Possessing Clinically Relevant Mutations. J Med Chem, 59, 6493-6500. PubMed id: 27308944

Date:

15-Mar-16 Release date: 28-Jun-17

Protein chain

P0A017  (DYR_STAAU) -  Dihydrofolate reductase from Staphylococcus aureus

Seq: 159 a.a.

Struc: 157 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: E.C.1.5.1.3 - dihydrofolate reductase.
• Pathway: Folate Coenzymes
• Reaction: (6S)-5,6,7,8-tetrahydrofolate + NADP⁺ = 7,8-dihydrofolate + NADPH + H⁺

(6S)-5,6,7,8-tetrahydrofolate + NADP(+) (Bound ligand (Het Group name = NAP) corresponds exactly) = 7,8-dihydrofolate + NADPH + H(+)

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

J Med Chem 59:6493-6500 (2016)

PubMed id: 27308944

Charged Propargyl-Linked Antifolates Reveal Mechanisms of Antifolate Resistance and Inhibit Trimethoprim-Resistant MRSA Strains Possessing Clinically Relevant Mutations.

S.M.Reeve, E.Scocchera, J.J.Ferreira, N.G-Dayanandan, S.Keshipeddy, D.L.Wright, A.C.Anderson.

ABSTRACT

Drug-resistant enzymes must balance catalytic function with inhibitor destabilization to provide a fitness advantage. This sensitive balance, often involving very subtle structural changes, must be achieved through a selection process involving a minimal number of eligible point mutations. As part of a program to design propargyl-linked antifolates (PLAs) against trimethoprim-resistant dihydrofolate reductase (DHFR) from Staphylococcus aureus, we have conducted a thorough study of several clinically observed chromosomal mutations in the enzyme at the cellular, biochemical, and structural levels. Through this work, we have identified a promising lead series that displays significantly greater activity against these mutant enzymes and strains than TMP. The best inhibitors have enzyme inhibition and MIC values near or below that of trimethoprim against wild-type S. aureus. Moreover, these studies employ a series of crystal structures of several mutant enzymes bound to the same inhibitor; analysis of the structures reveals a more detailed molecular understanding of drug resistance in this important enzyme.