Nature Structural Biology
3, 355 - 363 (1996)
doi:10.1038/nsb0496-355
Snapshot of an enzyme reaction intermediate in the structure of the ATP−Mg2+−oxalate ternary complex of Escherichia coli PEP carboxykinaseLeslie W. Tari1, Allan Matte1, Umarani Pugazhenthi1, Hughes Goldie2
& Louis T.J. Delbaere1
1Department of Biochemistry and, University of Saskatchewan, Saskatoon, 107 Wiggins Road Saskatchewan, Canada S7N5E5
2Department of Microbiology, University of Saskatchewan, Saskatoon, 107 Wiggins Road Saskatchewan, Canada S7N5E5 We report the 1.8 Å crystal structure of adenosine triphosphate (ATP)−magnesium−oxalate bound phosphoenolpyruvate carboxykinase (PCK) from Escherichia coli. ATP binding induces a 20° hinge-like rotation of the N- and C-terminal domains which closes the active-site cleft. PCK possesses a novel nucleotide-binding fold, particularly in the adenine-binding region, where the formation of a cis backbone torsion angle in a loop glycine residue promotes intimate contacts between the adenine-binding loop and adenine, while stabilizing a syn conformation of the base. This complex represents a reaction intermediate analogue along the pathway of the conversion of oxaloacetate to phosphoenolpyruvate, and provides insight into the mechanistic details of the chemical reaction catalysed by this enzyme.
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