Determination of Binding Stoichiometry by the Continuous Variation Method The Job Plot英文文献.pdfVIP

[27] C O N T I N U O U S V A R I A T I O N M E T H O D : T H E J OB P L O T 509 fluoride-phosphate pair and 0.2 for the D-amino acid oxidase inhibition by the riboflavin Y-sulfate-adenosine Y-sulfate pair were reported by Slater and Bonnet 2 and Yagi and Ozawa, z respectively. In the present LADH experiments, a values of 0.5 and 0.3 were obtained for the phen- anthroline-ADP and phenanthroline-AMP pairs, respectively. At pres- ent, it remains unexplained why such positive interactions occur between these pairs of inhibitors in the EIII2 complexes, but not between phen- anthroline and ADPR. The lack of interaction in the last case may be an occasional result of cancelling positive and negative interactions. The determination of a values for pairs between phenanthroline deriv- atives and ADPR derivatives may give useful information concerning the mechanism of these interactions as well as the distance between the Zn site and the ADPR-binding site of LADH. s9 It should be pointed out that the a value is closely related to the sta- bilization (= l/a) or destabilization (= a) factors operating between the coenzyme and inhibitor in ternary complexes of the enzyme. 1°For ex- ample, in the LADH-NADH-isobutyramide complex, the following re- lation was found: KE,NADH/gE,isobutyramide,NAD H = KE,isobutyramide/gE,NADn,isobutyramide = 60 = the stabilization factor = 1/a Thus ot = 0.016. 10 H. T h e o r e l l an d J. S. M c K i n l e y - M c K e e , Acta Chem. Scand. 15, 1797 (