1A95

XPRTASE FROM E. COLI COMPLEXED WITH MG:CPRPP AND GUANINE

Summary for 1A95

• Entry DOI: 10.2210/pdb1a95/pdb
• Descriptor: XANTHINE-GUANINE PHOSPHORIBOSYLTRANSFERASE, BORIC ACID, MAGNESIUM ION, ... (6 entities in total)
• Functional Keywords: phosphoribosyltransferase, transferase, purine salvage enzyme, glycosyltransferase
• Biological source: Escherichia coli
• Cellular location: Cell inner membrane; Peripheral membrane protein (Probable): P0A9M5
• Total number of polymer chains: 4
• Total formula weight: 69216.99

Authors

Vos, S.,Parry, R.J.,Burns, M.R.,De Jersey, J.,Martin, J.L. (deposition date: 1998-04-16, release date: 1998-11-11, Last modification date: 2024-05-22)

Primary citation

Vos, S.,Parry, R.J.,Burns, M.R.,de Jersey, J.,Martin, J.L.
Structures of free and complexed forms of Escherichia coli xanthine-guanine phosphoribosyltransferase.
J.Mol.Biol., 282:875-889, 1998

PubMed Abstract: Structures of free, substrate-bound and product-bound forms of Escherichia coli xanthine-guanine phosphoribosyltransferase (XGPRT) have been determined by X-ray crystallography. These are compared with the previously determined structure of magnesium and sulphate-bound XPRT. The structure of free XGPRT at 2.25 A resolution confirms the flexibility of residues in and around a mobile loop identified in other PRTases and shows that the cis-peptide conformation of Arg37 at the active site is maintained in the absence of bound ligands. The structures of XGPRT complexed with the purine base substrates guanine or xanthine in combination with cPRib-PP, an analog of the second substrate PRib-PP, have been solved to 2.0 A resolution. In these two structures the disordered phosphate-binding loop of uncomplexed XGPRT becomes ordered through interactions with the 5'-phosphate group of cPRib-PP. The cyclopentane ring of cPRib-PP has the C3 exo pucker conformation, stabilised by the cPRib-PP-bound Mg2+. The purine base specificity of XGPRT appears to be due to water-mediated interactions between the 2-exocyclic groups of guanine or xanthine and side-chains of Glu136 and Asp140, as well as the main-chain oxygen atom of Ile135. Asp92, together with Lys115, could help stabilise the N7-protonated tautomer of the incoming base and could act as a general base to remove the proton from N7 when the nucleotide product is formed. The 2.6 A resolution structure of XGPRT complexed with product GMP is similar to the substrate-bound complexes. However, the ribose ring of GMP is rotated by approximately 24 degrees compared with the equivalent ring in cPRib-PP. This rotation results in the loss of all interactions between the ribosyl group and the enzyme in the product complex.

PubMed: 9743633
DOI: 10.1006/jmbi.1998.2051

Experimental method

X-RAY DIFFRACTION (2 Å)