The 2.0 Å structure of malarial purine phosphoribosyltransferase in complex with a transition-state analogue inhibitor

Wuxian Shi, Caroline M. Li, Peter C. Tyler, Richard H. Furneaux, Sean M. Cahill, Mark E. Girvin, Charles Grubmeyer, Vern L. Schramm, Steven C. Almo

Research output: Contribution to journalArticle

128 Citations (Scopus)

Abstract

Malaria is a leading cause of worldwide mortality from infectious disease. Plasmodium falciparum proliferation in human erythrocytes requires purine salvage by hypoxanthine-guanine-xanthine phosphoribosyltransferase (HGXPRTase). The enzyme is a target for the development of novel antimalarials. Design and synthesis of transition-state analogue inhibitors permitted cocrystallization with the malarial enzyme and refinement of the complex to 2.0 Å resolution. Catalytic site contacts in the malarial enzyme are similar to those of human hypoxanthine-guanine phosphoribosyltransferase (HGPRTase) despite distinct substrate specificity. The crystal structure of malarial HGXPRTase with bound inhibitor, pyrophosphate, and two Mg2+ ions reveals features unique to the transition-state analogue complex. Substrate- assisted catalysis occurs by ribooxocarbenium stabilization from the O5' lone pair and a pyrophosphate oxygen. A dissociative reaction coordinate path is implicated in which the primary reaction coordinate motion is the ribosyl C1' in motion between relatively immobile purine base and (Mg)2-pyrophosphate. Several short hydrogen bonds form in the complex of the enzyme and inhibitor. The proton NMR spectrum of the transition-state analogue complex of malarial HGXPRTase contains two downfield signals at 14.3 and 15.3 ppm. Despite the structural similarity to the human enzyme, the NMR spectra of the complexes reveal differences in hydrogen bonding between the transition-state analogue complexes of the human and malarial HG(X)PRTases. The X-ray crystal structures and NMR spectra reveal chemical and structural features that suggest a strategy for the design of malaria-specific transition-state inhibitors.

Original languageEnglish (US)
Pages (from-to)9872-9880
Number of pages9
JournalBiochemistry
Volume38
Issue number31
DOIs
StatePublished - Aug 3 1999

Fingerprint

Nuclear magnetic resonance
Enzymes
Malaria
Hydrogen bonds
Crystal structure
Hypoxanthine Phosphoribosyltransferase
Salvaging
Antimalarials
Enzyme Inhibitors
Substrates
Hydrogen Bonding
Plasmodium falciparum
Substrate Specificity
Catalysis
Communicable Diseases
Protons
Hydrogen
Catalytic Domain
Stabilization
Erythrocytes

ASJC Scopus subject areas

  • Biochemistry

Cite this

The 2.0 Å structure of malarial purine phosphoribosyltransferase in complex with a transition-state analogue inhibitor. / Shi, Wuxian; Li, Caroline M.; Tyler, Peter C.; Furneaux, Richard H.; Cahill, Sean M.; Girvin, Mark E.; Grubmeyer, Charles; Schramm, Vern L.; Almo, Steven C.

In: Biochemistry, Vol. 38, No. 31, 03.08.1999, p. 9872-9880.

Research output: Contribution to journalArticle

Shi, Wuxian ; Li, Caroline M. ; Tyler, Peter C. ; Furneaux, Richard H. ; Cahill, Sean M. ; Girvin, Mark E. ; Grubmeyer, Charles ; Schramm, Vern L. ; Almo, Steven C. / The 2.0 Å structure of malarial purine phosphoribosyltransferase in complex with a transition-state analogue inhibitor. In: Biochemistry. 1999 ; Vol. 38, No. 31. pp. 9872-9880.
@article{de8d43a0725644208c78a67159819417,
title = "The 2.0 {\AA} structure of malarial purine phosphoribosyltransferase in complex with a transition-state analogue inhibitor",
abstract = "Malaria is a leading cause of worldwide mortality from infectious disease. Plasmodium falciparum proliferation in human erythrocytes requires purine salvage by hypoxanthine-guanine-xanthine phosphoribosyltransferase (HGXPRTase). The enzyme is a target for the development of novel antimalarials. Design and synthesis of transition-state analogue inhibitors permitted cocrystallization with the malarial enzyme and refinement of the complex to 2.0 {\AA} resolution. Catalytic site contacts in the malarial enzyme are similar to those of human hypoxanthine-guanine phosphoribosyltransferase (HGPRTase) despite distinct substrate specificity. The crystal structure of malarial HGXPRTase with bound inhibitor, pyrophosphate, and two Mg2+ ions reveals features unique to the transition-state analogue complex. Substrate- assisted catalysis occurs by ribooxocarbenium stabilization from the O5' lone pair and a pyrophosphate oxygen. A dissociative reaction coordinate path is implicated in which the primary reaction coordinate motion is the ribosyl C1' in motion between relatively immobile purine base and (Mg)2-pyrophosphate. Several short hydrogen bonds form in the complex of the enzyme and inhibitor. The proton NMR spectrum of the transition-state analogue complex of malarial HGXPRTase contains two downfield signals at 14.3 and 15.3 ppm. Despite the structural similarity to the human enzyme, the NMR spectra of the complexes reveal differences in hydrogen bonding between the transition-state analogue complexes of the human and malarial HG(X)PRTases. The X-ray crystal structures and NMR spectra reveal chemical and structural features that suggest a strategy for the design of malaria-specific transition-state inhibitors.",
author = "Wuxian Shi and Li, {Caroline M.} and Tyler, {Peter C.} and Furneaux, {Richard H.} and Cahill, {Sean M.} and Girvin, {Mark E.} and Charles Grubmeyer and Schramm, {Vern L.} and Almo, {Steven C.}",
year = "1999",
month = "8",
day = "3",
doi = "10.1021/bi990664p",
language = "English (US)",
volume = "38",
pages = "9872--9880",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "31",

}

TY - JOUR

T1 - The 2.0 Å structure of malarial purine phosphoribosyltransferase in complex with a transition-state analogue inhibitor

AU - Shi, Wuxian

AU - Li, Caroline M.

AU - Tyler, Peter C.

AU - Furneaux, Richard H.

AU - Cahill, Sean M.

AU - Girvin, Mark E.

AU - Grubmeyer, Charles

AU - Schramm, Vern L.

AU - Almo, Steven C.

PY - 1999/8/3

Y1 - 1999/8/3

N2 - Malaria is a leading cause of worldwide mortality from infectious disease. Plasmodium falciparum proliferation in human erythrocytes requires purine salvage by hypoxanthine-guanine-xanthine phosphoribosyltransferase (HGXPRTase). The enzyme is a target for the development of novel antimalarials. Design and synthesis of transition-state analogue inhibitors permitted cocrystallization with the malarial enzyme and refinement of the complex to 2.0 Å resolution. Catalytic site contacts in the malarial enzyme are similar to those of human hypoxanthine-guanine phosphoribosyltransferase (HGPRTase) despite distinct substrate specificity. The crystal structure of malarial HGXPRTase with bound inhibitor, pyrophosphate, and two Mg2+ ions reveals features unique to the transition-state analogue complex. Substrate- assisted catalysis occurs by ribooxocarbenium stabilization from the O5' lone pair and a pyrophosphate oxygen. A dissociative reaction coordinate path is implicated in which the primary reaction coordinate motion is the ribosyl C1' in motion between relatively immobile purine base and (Mg)2-pyrophosphate. Several short hydrogen bonds form in the complex of the enzyme and inhibitor. The proton NMR spectrum of the transition-state analogue complex of malarial HGXPRTase contains two downfield signals at 14.3 and 15.3 ppm. Despite the structural similarity to the human enzyme, the NMR spectra of the complexes reveal differences in hydrogen bonding between the transition-state analogue complexes of the human and malarial HG(X)PRTases. The X-ray crystal structures and NMR spectra reveal chemical and structural features that suggest a strategy for the design of malaria-specific transition-state inhibitors.

AB - Malaria is a leading cause of worldwide mortality from infectious disease. Plasmodium falciparum proliferation in human erythrocytes requires purine salvage by hypoxanthine-guanine-xanthine phosphoribosyltransferase (HGXPRTase). The enzyme is a target for the development of novel antimalarials. Design and synthesis of transition-state analogue inhibitors permitted cocrystallization with the malarial enzyme and refinement of the complex to 2.0 Å resolution. Catalytic site contacts in the malarial enzyme are similar to those of human hypoxanthine-guanine phosphoribosyltransferase (HGPRTase) despite distinct substrate specificity. The crystal structure of malarial HGXPRTase with bound inhibitor, pyrophosphate, and two Mg2+ ions reveals features unique to the transition-state analogue complex. Substrate- assisted catalysis occurs by ribooxocarbenium stabilization from the O5' lone pair and a pyrophosphate oxygen. A dissociative reaction coordinate path is implicated in which the primary reaction coordinate motion is the ribosyl C1' in motion between relatively immobile purine base and (Mg)2-pyrophosphate. Several short hydrogen bonds form in the complex of the enzyme and inhibitor. The proton NMR spectrum of the transition-state analogue complex of malarial HGXPRTase contains two downfield signals at 14.3 and 15.3 ppm. Despite the structural similarity to the human enzyme, the NMR spectra of the complexes reveal differences in hydrogen bonding between the transition-state analogue complexes of the human and malarial HG(X)PRTases. The X-ray crystal structures and NMR spectra reveal chemical and structural features that suggest a strategy for the design of malaria-specific transition-state inhibitors.

UR - http://www.scopus.com/inward/record.url?scp=0033519996&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033519996&partnerID=8YFLogxK

U2 - 10.1021/bi990664p

DO - 10.1021/bi990664p

M3 - Article

C2 - 10433693

AN - SCOPUS:0033519996

VL - 38

SP - 9872

EP - 9880

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 31

ER -