PDBsum entry 1g6c

Transferase

PDB id: 1g6c

Contents

Protein chains

226 a.a. *

Ligands

IFP ×2

POP ×2

TZP ×2

Metals

_MG ×2

Waters ×293

* Residue conservation analysis

References listed in PDB file

Key reference

• Title: Structural characterization of the enzyme-Substrate, Enzyme-Intermediate, And enzyme-Product complexes of thiamin phosphate synthase.
• Authors: D.H.Peapus, H.J.Chiu, N.Campobasso, J.J.Reddick, T.P.Begley, S.E.Ealick.
• Ref.: Biochemistry, 2001, 40, 10103-10114. [DOI no: 10.1021/bi0104726]
• PubMed id: 11513589

Abstract

Thiamin phosphate synthase catalyzes the formation of thiamin phosphate from 4-amino-5-(hydroxymethyl)-2-methylpyrimidine pyrophosphate and 5-(hydroxyethyl)-4-methylthiazole phosphate. Several lines of evidence suggest that the reaction proceeds via a dissociative mechanism. The previously determined crystal structure of thiamin phosphate synthase in complex with the reaction products, thiamin phosphate and magnesium pyrophosphate, provided a view of the active site and suggested a number of additional experiments. We report here seven new crystal structures primarily involving crystals of S130A thiamin phosphate synthase soaked in solutions containing substrates or products. We prepared S130A thiamin phosphate synthase with the intent of characterizing the enzyme-substrate complex. Surprisingly, in three thiamin phosphate synthase structures, the active site density cannot be modeled as either substrates or products. For these structures, the best fit to the electron density is provided by a model that consists of independent pyrimidine, pyrophosphate, and thiazole phosphate fragments, consistent with a carbenium ion intermediate. The resulting carbenium ion is likely to be further stabilized by proton transfer from the pyrimidine amino group to the pyrophosphate to give the pyrimidine iminemethide, which we believe is the species that is observed in the crystal structures.

Secondary reference #1

• Title: Mechanistic studies on thiamin phosphate synthase: evidence for a dissociative mechanism.
• Authors: J.J.Reddick, R.Nicewonger, T.P.Begley.
• Ref.: Biochemistry, 2001, 40, 10095-10102. [DOI no: 10.1021/bi010267q]
• PubMed id: 11513588

Secondary reference #2

• Title: Crystal structure of thiamin phosphate synthase from bacillus subtilis at 1.25 a resolution.
• Authors: H.J.Chiu, J.J.Reddick, T.P.Begley, S.E.Ealick.
• Ref.: Biochemistry, 1999, 38, 6460-6470. [DOI no: 10.1021/bi982903z]
• PubMed id: 10350464

Secondary reference #3

• Title: Characterization of the bacillus subtilis thic operon involved in thiamine biosynthesis.
• Authors: Y.Zhang, S.V.Taylor, H.J.Chiu, T.P.Begley.
• Ref.: J Bacteriol, 1997, 179, 3030-3035.
• PubMed id: 9139923