Using experimental evolution to probe molecular mechanisms of protein function.
Directed evolution is a powerful tool for engineering protein function. The process of directed evolution involves iterative rounds of sequence diversification followed by assaying activity of variants and selection. The range of sequence variants and linked activities generated in the course of an evolution are a rich information source for investigating relationships between sequence and function. Key residue positions determining protein function, combinatorial contributors to activity and even potential functional mechanisms have been revealed in directed evolutions. The recent application of high throughput sequencing substantially increases the information that can be retrieved from directed evolution experiments. Combined with computational analysis this additional sequence information has allowed high-resolution analysis of individual residue contributions to activity. These developments promise to significantly enhance the depth of insight that experimental evolution provides into mechanisms of protein function.
Department of Molecular and Cell Biology, University of Leicester, Henry Wellcome Building, Lancaster Road, Leicester, LE1 9HN, United Kingdom.|Department of Cardiovascular Sciences, University of Leicester, Henry Wellcome Building, Lancaster Road, Leicester, LE1 9HN, United Kingdom.
Marlies
M
Fischer
Department of Molecular and Cell Biology, University of Leicester, Henry Wellcome Building, Lancaster Road, Leicester, LE1 9HN, United Kingdom.|Department of Cardiovascular Sciences, University of Leicester, Henry Wellcome Building, Lancaster Road, Leicester, LE1 9HN, United Kingdom.
Mandeep
M
Kang
Department of Molecular and Cell Biology, University of Leicester, Henry Wellcome Building, Lancaster Road, Leicester, LE1 9HN, United Kingdom.|Department of Cardiovascular Sciences, University of Leicester, Henry Wellcome Building, Lancaster Road, Leicester, LE1 9HN, United Kingdom.
Nicholas Pj
NP
Brindle
D000818:Animals; D019020:Directed Molecular Evolution / Q000379:methods; D006801:Humans; D009154:Mutation; D015202:Protein Engineering / Q000379:methods; D011506:Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D020539:Sequence Analysis, Protein / Q000379:methods
D011506:Proteins
directed evolution; evolution; high throughput sequencing; mutation; next generation sequencing; protein function; protein sequence