Leishmania mexicana mexicana glucose-6-phosphate isomerase: crystallization, molecular-replacement solution and inhibition.
Glucose-6-phosphate isomerase (PGI; EC 5.3.1.9; also often called by its old nomenclature phosphoglucose isomerase) is an intracellular enzyme that catalyses the reversible conversion of D-glucose 6-phosphate (G6P) to D-fructose 6-phosphate (F6P). The native Leishmania PGI is a homodimeric molecule of 60 kDa per monomer with 47% sequence identity to human PGI. It has been shown to be present in both the cytosol and the glycosome of Leishmania promastigotes and represents a potential target for rational drug design. The present work describes the crystallization of two bacterially expressed Leishmania PGI constructs, one corresponding to the natural protein and the other to an N-terminally deleted form. Crystals of both forms are identical and present a large c unit-cell parameter. A complete data set was collected from the N-terminally deleted PGI to a resolution of 3.3 A in space group P6(1), with unit-cell parameters a = b = 87.0, c = 354.7 A, alpha = beta = 90, gamma = 120 degrees. A preliminary study of the first inhibitors to be evaluated on the Leishmania enzyme is also reported.
Cordeiro
Artur T
AT
Laboratory of Protein Crystallography and Structural Biology, Physics Institute of São Carlos, University of São Paulo, USP, Av. Trabalhador Sãocarlense 400, PO Box 369, 13566-590 São Carlos-SP, Brazil.
Hardré
Renaud
R
Michels
Paul A M
PA
Salmon
Laurent
L
Delboni
Luis F
LF
Thiemann
Otavio H
OH
D000595:Amino Acid Sequence; D000818:Animals; D003460:Crystallization; D018360:Crystallography, X-Ray; D019281:Dimerization; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005636:Fructosephosphates / Q000378:metabolism; D005956:Glucose-6-Phosphate Isomerase / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D020128:Inhibitory Concentration 50; D007700:Kinetics; D007894:Leishmania mexicana / Q000201:enzymology*; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment
D004791:Enzyme Inhibitors; D005636:Fructosephosphates; D011994:Recombinant Proteins; C027618:fructose-6-phosphate; D005956:Glucose-6-Phosphate Isomerase