《丙氨酸扫描》.pdfVIP

302 Combinatorial alanine-scanning Kim L Morrison and Gregory A Weiss* Combinatorial libraries of alanine-substituted proteins can be oligonucleotide-directed mutagenesis to measure used to rapidly identify residues important for protein function, hydrogen-bond strengths in proteins. stability and shape. Each alanine substitution examines the contribution of an individual amino acid sidechain to the Combinatorial libraries of mutant proteins can be used to functionality of the protein. The recently described method of rapidly analyze protein function and identify important shotgun scanning uses phage-displayed libraries of alanine- sidechain functionalities. For the examination of recep- substituted proteins for high-throughput analysis. tor–ligand interactions, protein libraries can be generated with either random [3,4,5•] or site-specific mutations. Addresses Saturation mutagenesis substitutes specific positions with Department of Chemistry, University of California, Irvine, all 20 naturally occurring amino acids [6,7]. However, sub- CA 92697-2025, USA stitution of a few carefully chosen amino acids can provide *e-mail: gweiss@uci.edu a thorough portrait of protein function and simplify analy- Current Opinion in Chemical Biology 2001, 5:302–307 sis of the data. This review focuses upon combinatorial protein libraries substituted with either alanine or the 1367-5931/01/$ — see front