pH-Dependent proteolytic activity of histidine-pendant polyacrylamides

Artificial enzymes with serine, aspartic acid, and histidine that mimic the catalytic triad of serine proteases have been studied. However, they were mainly investigated for their cleavage activity on low molecular-weight esters and amides, and there has been a lack of knowledge about their protein cleavage activity. We previously found that not only ternary polyacrylamides with serine, aspartic acid, and histidine but also homo-polyacrylamides with a single type of amino acid cleaved bovine and human serum albumins. These results suggested that the carboxy group, a common functional group in active polymers, may be involved in the expression of activity. In this study, we synthesized histidine polymers in which the carboxy group was changed to a carboxamide or amino group and investigated the relationships between structure and activity. Histidine homopolymers and histidine-acrylamide copolymers were incubated with bovine serum albumin at 37°C for one week under different pH conditions and then analyzed by sodium dodecyl-sulfate polyacrylamide gel electrophoresis. In the case of histidine homopolymers, the carboxy-type homopolymers were active under weakly acidic to neutral conditions, while the carboxamido- and amino-type homopolymers were not active regardless of liquid properties. In the case of histidine-acrylamide copolymers, their activity was essentially the same as that of each type of histidine homopolymer, but for all types of copolymers, activity appeared under weakly basic conditions when the histidine monomer units were of a specific density. The carboxy-type copolymers with dual-mode activity showed different degradation patterns of bovine serum albumin under acidic and basic conditions. The structural features common to the active homopolymers and copolymers suggested that the carboxy group contributes to the activity under weakly acidic to neutral conditions, while the imidazole group of the histidine side chain and its density contribute to the activity under weakly basic conditions.