Crystallographic structure of allosterically inhibited phosphofructokinase at 7 A resolution.

The structure of the allosterically inhibited form of phosphofructokinase from Bacillus stearothermophilus has been determined by X-ray crystallography to 7 A resolution by molecular replacement using the known structure of the active state as a starting model. Comparing the inhibited state with the active state, the tetramer is twisted about its long axis such that one pair of subunits in the tetramer rotates relative to the other pair by about 8 degrees around one of the molecular dyad axes. This rotation partly closes the binding site for the co-operative substrate fructose-6-phosphate, explaining its weaker binding to this conformational state. Within the subunit, one domain rotates relative to the other by 4.5 degrees, which further closes the fructose-6-phosphate site, without closing the cleft between the domains of the same subunit: this motion causes little change to the catalytic site. This T-state model is consistent with the simple allosteric kinetic scheme in which the active and the inhibited conformations differ in their affinities for fructose-6-phosphate, but not in their catalytic rates. It does not explain the heterotropic allosteric effects.