Crystal structure of Trypanosoma cruzi glyceraldehyde-3-phosphate dehydrogenase complexed with an analogue of 1,3-bisphospho-d-glyceric acid.
We report here the first crystal structure of a stable isosteric analogue of 1,3-bisphospho-d-glyceric acid (1,3-BPGA) bound to the catalytic domain of Trypanosoma cruzi glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) in which the two phosphoryl moieties interact with Arg249. This complex possibly illustrates a step of the catalytic process by which Arg249 may induce compression of the product formed, allowing its expulsion from the active site. Structural modifications were introduced into this isosteric analogue and the respective inhibitory effects of the resulting diphosphorylated compounds on T. cruzi and Trypanosoma brucei gGAPDHs were investigated by enzymatic inhibition studies, fluorescence spectroscopy, site-directed mutagenesis, and molecular modelling. Despite the high homology between the two trypanomastid gGAPDHs (> 95%), we have identified specific interactions that could be used to design selective irreversible inhibitors against T. cruzi gGAPDH.
Ladame
Sylvain
S
Laboratoire de Synthèse et de Physico-Chimie de Molécules d'Intérêt Biologique UMR-CNRS 5068, Université Paul Sabatier, Toulouse, France. sl324@cam.ac.uk
Castilho
Marcelo S
MS
Silva
Carlos H T P
CH
Denier
Colette
C
Hannaert
Véronique
V
Périé
Jacques
J
Oliva
Glaucius
G
Willson
Michèle
M
D000818:Animals; D001665:Binding Sites; D018360:Crystallography, X-Ray; D004163:Diphosphoglyceric Acids / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+) / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism*; D007700:Kinetics; D008958:Models, Molecular; D008968:Molecular Conformation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D013045:Species Specificity; D055672:Static Electricity; D013329:Structure-Activity Relationship; D014346:Trypanosoma brucei brucei / Q000201:enzymology; D014349:Trypanosoma cruzi / Q000201:enzymology*
D004163:Diphosphoglyceric Acids; D004791:Enzyme Inhibitors; C015891:glycerate 1,3-biphosphate; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+)