By Jaroslava Turková (Eds.)
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Additional info for Affinity Chromatography
10) and the experimental binding isotherm did not tend to zero abruptly (Fig. 8). This inconsistency can be interpreted as follows. As the ratio of cu to c is 102-103 and the amount of diphosphate in polyuridylic acid is 102-103, the loop formation of poly- QUANTITATIVE EVALUATION OF SPECIFIC COMPLEXES 46 uridylic acid with oligoadenylic acid as a clasp can be neglected (although it cannot be neglected for infinitely long polymers). Hence loops can be neglected in double- or triplestranded helices.
95-178. H. , Chromarographia, 4 (1971) 578-581. , Cuatrecasas, P. , J. Bwl. , 246 (1971) 196-200. , Coupek, J. ,Eiochim. Biophys, Acta, 427 (1976) 586-593. 35 Chapter 4 Application of affinity chromatography to the quantitative evaluation of specific complexes Association processes undoubtedly play one of the most important roles in biochemical processes. The initial degree of bonding to a substrate has an important effect on the catalytic properties of an enzyme. Control processes in biology depend on the association of the repressor with the operon, hormones with specific receptors, etc.
Under experimental conditions, the complex of polyuridylic acid with oligoadenylic acid is in a triple-stranded helical state when in solution. Therefore, when the binding isotherm is calculated, the loop entropy of two strands of polyuridylic acid was taken into consideration. 16) where A = 410g(l - 4 p ) -k 4Li2(dp) i- p In the case when p is taken as a parameter, the binding isotherm can be obtained. If the excluded volume effect is taken into consideration, the parameter a can be determined as 2, which appears in a factor due to loops in the partition function, 2n~b(j+l)/(j+2)a.