1TYS

WATER-MEDIATED SUBSTRATE(SLASH)PRODUCT DISCRIMINATION: THE PRODUCT COMPLEX OF THYMIDYLATE SYNTHASE AT 1.83 ANGSTROMS

Summary for 1TYS

• Entry DOI: 10.2210/pdb1tys/pdb
• Descriptor: THYMIDYLATE SYNTHASE, THYMIDINE-5'-PHOSPHATE, DIHYDROFOLIC ACID, ... (4 entities in total)
• Functional Keywords: transferase, methyltransferase
• Biological source: Escherichia coli
• Cellular location: Cytoplasm: P00470
• Total number of polymer chains: 1
• Total formula weight: 31309.22

Authors

Fauman, E.,Rutenber, E.,Stroud, R. (deposition date: 1993-05-07, release date: 1994-06-22, Last modification date: 2024-11-20)

Primary citation

Fauman, E.B.,Rutenber, E.E.,Maley, G.F.,Maley, F.,Stroud, R.M.
Water-mediated substrate/product discrimination: the product complex of thymidylate synthase at 1.83 A.
Biochemistry, 33:1502-1511, 1994

PubMed Abstract: In an irreversible enzyme-catalyzed reaction, strong binding of the products would lead to substantial product inhibition. The X-ray crystal structure of the product complex of thymidylate synthase (1.83-A resolution, R factor = 0.183 for all data between 7.0 and 1.83 A) identifies a bound water molecule that serves to disfavor binding of the product nucleotide, dTMP. This water molecule is hydrogen bonded to absolutely conserved Tyr 146 (using the Lactobacillus casei numbering system) and is displaced by the C7 methyl group of the reaction product thymidylate. The relation between this observation and kinetic and thermodynamic values is discussed. The structure reveals a carbamate modified N-terminus that binds in a highly conserved site, replaced by side chains that can exploit the same site in other TS sequences. The enzyme-products complex is compared to the previously determined structure of enzyme-substrate-cofactor analog. This comparison reveals changes that occur between the first covalent complex formed between enzyme and substrate with an inhibitory cofactor analog and the completed reaction. The almost identical arrangement of ligands in these two structures contributes to our model for the TS reaction and verifies the physiological relevance of the mode in which potent inhibitors bind to this target for rational drug design.

PubMed: 8312270
DOI: 10.1021/bi00172a029

Experimental method

X-RAY DIFFRACTION (1.8 Å)