INCREASED ACTIVITY OF PORCINE PANCREATIC PHOSPHOLIPASE A(2) BY DESIGNED LONG-RANGE ELECTROSTATIC STABILIZATION OF THE TRANSITION-STATE

Stabilisation of the catalytic transition state by long-range charge i nteractions has been tested with mutagenesis for porcine pancreatic ph ospholipase A(2). Electrostatics calculations were used to determine l ocations which would interact preferentially with one part of the dipo lar charge separation that is believed to develop in the transition st ate. Experiment shows increased enzyme activity relative to wild-type recombinant enzyme for mutants N97D and N101D, consistent with the des ign. (C) 1995 Academic Press, Inc.