Combinatorial synthesis, selection, and properties of esterase peptide dendrimers.
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) approximately 0.1 mM, catalytic rate constants k(cat) approximately 0.1 min(-1), and specific rate accelerations over background up to k(cat)/k(uncat) = 10,000.
Clouet
Anthony
A
Department of Chemistry and Biochemistry, University of Berne, Freiestrasse 3, CH-3012 Berne, Switzerland.
Darbre
Tamis
T
Reymond
Jean-Louis
JL
D001190:Arylsulfonates / Q000737:chemistry; D020650:Combinatorial Chemistry Techniques; D050091:Dendrimers / Q000138:chemical synthesis* / Q000737:chemistry; D004950:Esterases / Q000138:chemical synthesis* / Q000737:chemistry; D006868:Hydrolysis; D007700:Kinetics; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry
D001190:Arylsulfonates; D050091:Dendrimers; D010455:Peptides; D004950:Esterases