Combinatorial synthesis, selection, and properties of esterase peptide dendrimers

Abstract

Abstract A 65,536‐member combinatorial library of peptide dendrimers was prepared by split‐and‐mix synthesis and screened on solid support for esterolytic activity in aqueous buffer using 8‐butyryloxypyrene‐1,3,6‐trisulfonate ( 2 ) as a fluorogenic substrate. Active sequences were identified by analysis of fluorescent beads. The corresponding dendrimers were resynthesized by solid‐phase synthesis, cleaved from the resin, and purified by preparative reverse‐phase HPLC. The dendrimers showed the expected catalytic activity in aqueous buffer. Catalysis was studied against a pannel of fluorogenic 8‐acyloxypyrene‐1,3,6‐trisulfonate substrates. The catalytic peptide dendrimers display enzyme‐like kinetics in aqueous buffer with substrate binding in the range K M ˜ 0.1 m M , catalytic rate constants k cat ˜ 0.1 min –1 , and specific rate accelerations over background up to k cat / k uncat = 10,000. © 2005 Wiley Periodicals, Inc. Biopolymers 84: 114–123, 2006 This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com