Crystal structure of phosphoglucose isomerase from Trypanosoma brucei complexed with glucose-6-phosphate at 1.6 A resolution.
Enzymes of glycolysis in Trypanosoma brucei have been identified as potential drug targets for African sleeping sickness because glycolysis is the only source of ATP for the bloodstream form of this parasite. Several inhibitors were previously reported to bind preferentially to trypanosomal phosphoglucose isomerase (PGI, the second enzyme in glycolysis) than to mammalian PGIs, which suggests that PGI might make a good target for species-specific drug design. Herein, we report recombinant expression, purification, crystallization and X-ray crystal structure determination of T. brucei PGI. One structure solved at 1.6 A resolution contains a substrate, D-glucose-6-phosphate, in an extended conformation in the active site. A second structure solved at 1.9 A resolution contains a citrate molecule in the active site. The structures are compared with the crystal structures of PGI from humans and from Leishmania mexicana. The availability of recombinant tPGI and its first high-resolution crystal structures are initial steps in considering this enzyme as a potential drug target.
Laboratory for Molecular Biology, Department of Biological Sciences, University of Illinois, Chicago, Illinois 60607, USA.
Diana
D
Arsenieva
Brian L
BL
Appavu
Gloria H
GH
Mazock
Constance J
CJ
Jeffery
D000818:Animals; D001665:Binding Sites; D019343:Citric Acid / Q000737:chemistry; D018360:Crystallography, X-Ray; D019298:Glucose-6-Phosphate / Q000737:chemistry*; D005956:Glucose-6-Phosphate Isomerase / Q000737:chemistry* / Q000302:isolation & purification; D006801:Humans; D007894:Leishmania mexicana / Q000201:enzymology; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D014346:Trypanosoma brucei brucei / Q000201:enzymology*
D011994:Recombinant Proteins; D019343:Citric Acid; D019298:Glucose-6-Phosphate; D005956:Glucose-6-Phosphate Isomerase