85 



-9 

 polymerase and possessed an apparent K of 6.9 x 10 M. 



The inhibition seen with ADGG-BSA clearly deviated from the 

 strictly noncompetitive, as was the case with ADH-BSA con- 

 jugates. An average of the three x-intercepts resulted in 

 an apparent K for ADGG-BSA of 127 x 10 M. 



These studies verified the retention by ADGG of the 

 inhibitory potential of free a-amanitin and demonstrated 

 that its carbodiimide mediated conjugation to proteins was 

 feasible. From the previously described attempts at conju- 

 gation of ADH and Con A, the need for a system to evaluate 

 the conditions under which successful conjugation of ADGG 

 to Con A could be obtained became apparent. The optimal 

 conditions for production of conjugate that retained both 

 a-amanitin and lectin associated properties required defini- 

 tion. For this reason and to conserve ADGG, the kinetic 



14 

 analysis of the EDC mediated binding of " C-hippuric acid 



and Con A was undertaken. 



Hippuric Acid-Con A Conjugates 



Synthesis 



The conditions which would allow for the introduction 



14 

 of a defined number of ' C-labeled hippuric acid residues 



onto Con A with a minimum of protein cross-linking were 

 determined by measuring the amount of C activity preci- 

 pitated by TCA and collected on GF/C glass fiber filters as 

 described in Materials and Methods. Figure 16 presents the 

 results of an experiment in which the effect of varying pH 



