1DQP

CRYSTAL STRUCTURE OF GIARDIA GUANINE PHOSPHORIBOSYLTRANSFERASE COMPLEXED WITH IMMUCILLING

Summary for 1DQP

• Entry DOI: 10.2210/pdb1dqp/pdb
• Related: 1DQN
• Descriptor: GUANINE PHOSPHORIBOSYLTRANSFERASE, 1,4-DIDEOXY-1,4-IMINO-1-(S)-(9-DEAZAGUANIN-9-YL)-D-RIBITOL, ISOPROPYL ALCOHOL, ... (4 entities in total)
• Functional Keywords: protein-inhibitor complex, immucilling, 9-deazaguanine, transferase
• Biological source: Giardia intestinalis
• Total number of polymer chains: 2
• Total formula weight: 53417.28

Authors

Shi, W.,Munagala, N.R.,Wang, C.C.,Li, C.M.,Tyler, P.C.,Furneaux, R.H.,Grubmeyer, C.,Schramm, V.L.,Almo, S.C. (deposition date: 2000-01-04, release date: 2000-07-26, Last modification date: 2024-02-07)

Primary citation

Shi, W.,Munagala, N.R.,Wang, C.C.,Li, C.M.,Tyler, P.C.,Furneaux, R.H.,Grubmeyer, C.,Schramm, V.L.,Almo, S.C.
Crystal structures of Giardia lamblia guanine phosphoribosyltransferase at 1.75 A(,).
Biochemistry, 39:6781-6790, 2000

PubMed Abstract: Giardia lamblia, the protozoan parasite responsible for giardiasis, requires purine salvage from its host for RNA and DNA synthesis. G. lamblia expresses an unusual purine phosphoribosyltransferase with a high specificity for guanine (GPRTase). The enzyme's sequence significantly diverges from those of related enzymes in other organisms. The transition state analogue immucillinGP is a powerful inhibitor of HGXPRTase from malaria [Li, C. M., et al. (1999) Nat. Struct. Biol. 6, 582-587] and is also a 10 nM inhibitor of G. lamblia GPRTase. Cocrystallization of GPRTase with immucillinGP led unexpectedly to a GPRTase.immucillinG binary complex with an open catalytic site loop. Diffusion of ligands into preformed crystals gave a GPRTase.immucillinGP.Mg(2+).pyrophosphate complex in which the open loop is stabilized by crystal contacts. G. lamblia GPRTase exhibits substantial structural differences from known purine phosphoribosyltransferases at positions remote from the catalytic site, but conserves most contacts to the bound inhibitor. The filled catalytic site with an open catalytic loop provides insight into ligand binding. One active site Mg(2+) ion is chelated to pyrophosphate, but the other is chelated to two conserved catalytic site carboxylates, suggesting a role for these amino acids. This arrangement of Mg(2+) and pyrophosphate has not been reported in purine phosphoribosyltransferases. ImmucillinG in the binary complex is anchored by its 9-deazaguanine group, and the iminoribitol is disordered. No Mg(2+) or pyrophosphate is detected; thus, the 5'-phosphoryl group is needed to immobilize the iminoribitol prior to magnesium pyrophosphate binding. Filling the catalytic site involves (1) binding the purine ring, (2) anchoring the 5'-phosphate to fix the ribosyl group, (3) binding the first Mg(2+) to Asp125 and Glu126 carboxyl groups and binding Mg(2+).pyrophosphate, and (4) closing the catalytic site loop and formation of bound (Mg(2+))(2). pyrophosphate prior to catalysis. Guanine specificity is provided by two peptide carbonyl oxygens hydrogen-bonded to the exocyclic amino group and a weak interaction to O6. Transition state formation involves N7 protonation by Asp129 acting as the general acid.

PubMed: 10841757
DOI: 10.1021/bi000128t

Experimental method

X-RAY DIFFRACTION (1.75 Å)