Modeling the inhibitor activity and relative binding affinities in enzyme-inhibitor-protein systems: application to developmental regulation in a PG-PGIP system.
Within a number of classes of hydrolytic enzymes are certain enzymes whose activity is modulated by a specific inhibitor-protein that binds to the enzyme and forms an inactive complex. One unit of a specific inhibitor-protein activity is often defined as the amount necessary to inhibit one unit of its target enzyme by 50 %. No objective quantitative means is available to determine this point of 50 % inhibition in crude systems such as those encountered during purification. Two models were derived: the first model is based on an irreversible binding approximation, and the second, or equilibrium, model is based on reversible binding. The two models were validated using the inhibition data for the polygalacturonase-polygalacturonase-inhibiting protein (PG-PGIP) system. Theory and experimental results indicate that the first model can be used for inhibitor protein activity determination and the second model can be used for inhibitor protein activity determination as well as for comparison of association constants among enzymes and their inhibitor-proteins from multiple sources. The models were used to identify and further clarify the nature of a differential regulation of expression of polygalacturonase-inhibiting protein in developing cantaloupe fruit. These are the first relations that provide for an objective and quantitative determination of inhibitor-protein activity in both pure and crude systems. Application of these models should prove valuable in gaining insights into regulatory mechanisms and enzyme-inhibitor-protein interactions.
Fish
Wayne W
WW
USDA-ARS, SCARL, P.O. Box 159, Lane, Oklahoma 74555, USA. wfish-usda@lane-ag.org
Madihally
Sundararajan V
SV
D003198:Computer Simulation; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000737:chemistry; D004795:Enzyme Stability; D007700:Kinetics; D008956:Models, Chemical; D046912:Multiprotein Complexes / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry*; D011096:Polygalacturonase / Q000037:antagonists & inhibitors / Q000737:chemistry*; D011485:Protein Binding
D004791:Enzyme Inhibitors; D046912:Multiprotein Complexes; C081479:PGIP protein, plant; D010940:Plant Proteins; D011096:Polygalacturonase