Reactions at prochiral centers. Interdependence in the estimation of enzyme stereospecificity toward prochiral centers and the configurational purity of labeled substrates.
In many enzymic reactions one or the other hydrogen is released from a prochiral center abCH2. To determine the stereospecificity with respect to such centers under nonreversal conditions a substrate with a chiral center abC1HH is employed (H = 3H or 2H). Assuming the simplest possible model, the R specificity of an enzyme or enzyme system (eR) may be defined in terms of the configurational R purity of the substrate with respect to the labeled center (pR), and the fraction of H released (r) or conserved (l - r) during the reaction by the equation eR = [(l - r) -pR]/[1 - 2pR]. Where r is constant, this is a rectangular hyperbola. The apparent stereospecificity calculated with this equation serves as a characteristic of the experimental system irrespective of the assumed model. The problems arising from direct isotope effects are discussed and the theoretical advantages of using product 3H:14C ratios under low conversion conditions noted. Applications of the equation to the determination of configurational purity at labeled centers, to the study of stereospecificity in enzymes and enzyme systems, and to the study of the biosynthesis of natural products are discussed.
D001665:Binding Sites; D004798:Enzymes / Q000378:metabolism*; D007700:Kinetics; D008433:Mathematics; D008968:Molecular Conformation; D011485:Protein Binding; D013237:Stereoisomerism
D004798:Enzymes