Polyanionic inhibitors of phosphoglycerate mutase: combined structural and biochemical analysis.
The effects that the inhibitors inositol hexakisphosphate and benzene tri-, tetra- and hexacarboxylates have on the phosphoglycerate mutases from Saccharomyces cerevisiae and Schizosaccharomyces pombe have been determined. Their Kivalues have been calculated, and the ability of the inhibitors to protect the enzymes against limited proteolysis investigated. These biochemical data have been placed in a structural context by the solution of the crystal structures of S. cerevisiae phosphoglycerate mutase soaked with inositol hexakisphosphate or benzene hexacarboxylate. These large polyanionic compounds bind to the enzyme so as to block the entrance to the active-site cleft. They form multiple interactions with the enzyme, consistent with their low Kivalues, and afford good protection against limited proteolysis of the C-terminal region by thermolysin. The inositol compound is more efficacious because of its greater number of negative charges. The S. pombe phosphoglycerate mutase that is inherently lacking a comparable C-terminal region has higher Kivalues for the compounds tested. Moreover, the S. pombe enzyme is less sensititive to proteolysis, and the presence or absence of the inhibitor molecules has little effect on susceptibility to proteolysis.
Rigden
D J
DJ
School of Biochemistry and Molecular Biology, University of Leeds, Leeds, LS2 9JT, England.
Walter
R A
RA
Phillips
S E
SE
Fothergill-Gilmore
L A
LA
D000595:Amino Acid Sequence; D001555:Benzene Derivatives / Q000378:metabolism*; D018360:Crystallography, X-Ray; D004791:Enzyme Inhibitors / Q000378:metabolism*; D008024:Ligands; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010736:Phosphoglycerate Mutase / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D010833:Phytic Acid / Q000378:metabolism*; D011487:Protein Conformation; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D012568:Schizosaccharomyces / Q000201:enzymology*; D013820:Thermolysin
D001555:Benzene Derivatives; D004791:Enzyme Inhibitors; D008024:Ligands; D010833:Phytic Acid; D013820:Thermolysin; D010736:Phosphoglycerate Mutase