1TSX

THYMIDYLATE SYNTHASE R179E MUTANT

Summary for 1TSX

• Entry DOI: 10.2210/pdb1tsx/pdb
• Descriptor: THYMIDYLATE SYNTHASE, 2'-DEOXYURIDINE 5'-MONOPHOSPHATE (3 entities in total)
• Functional Keywords: transferase, methyltransferase, nucleotide biosynthesis
• Biological source: Lactobacillus casei
• Cellular location: Cytoplasm: P00469
• Total number of polymer chains: 1
• Total formula weight: 36910.56

Authors

Finer-Moore, J.,Stroud, R.M. (deposition date: 1995-12-05, release date: 1996-03-08, Last modification date: 2024-02-14)

Primary citation

Finer-Moore, J.S.,Fauman, E.B.,Morse, R.J.,Santi, D.V.,Stroud, R.M.
Contribution of a salt bridge to binding affinity and dUMP orientation to catalytic rate: mutation of a substrate-binding arginine in thymidylate synthase.
Protein Eng., 9:69-75, 1996

PubMed Abstract: Invariant arginine 179, one of four arginines that are conserved in all thymidylate synthases (TS) and that bind the phosphate moiety of the substrate 2'-deoxyuridine-5'-monophosphate (dUMP), can be altered even to a negatively charged glutamic acid with little effect on kcat. In the mutant structures, ordered water or the other phosphate-binding arginines compensate for the hydrogen bonds made by Arg179 in the wild-type enzyme and there is almost no change in the conformation or binding site of dUMP. Correlation of dUMP Kds for TS R179A and TS R179K with the structures of their binary complexes shows, that the positive charge on Arg179 contributes significantly to dUMP binding affinity. kcat/K(m) for dUMP measures the rate of dUMP binding to TS during the ordered bi-substrate reaction, and in the ternary complex dUMP provides a binding surface for the cofactor. kcat/K(m) reflects the ability of the enzyme to accept a properly oriented dUMP for catalysis and is less sensitive than is Kd to the changes in electrostatics at the phosphate binding site.

PubMed: 9053905
DOI: 10.1093/protein/9.1.69

Experimental method

X-RAY DIFFRACTION (2.5 Å)