Más notícias para Darwin: cada posição tem características únicas que não podem ser capturadas por algumas mutações

domingo, fevereiro 06, 2011

High-resolution mapping of protein sequence-function relationships

Douglas M Fowler, Carlos L Araya, Sarel J Fleishman, Elizabeth H Kellogg, Jason J Stephany, David Baker & Stanley Fields

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Nature Methods 7, 741–746 (2010) doi:10.1038/nmeth.1492

Received 25 March 2010 Accepted 13 July 2010 Published online 15 August 2010

Abstract

We present a large-scale approach to investigate the functional consequences of sequence variation in a protein. The approach entails the display of hundreds of thousands of protein variants, moderate selection for activity and high-throughput DNA sequencing to quantify the performance of each variant. Using this strategy, we tracked the performance of >600,000 variants of a human WW domain after three and six rounds of selection by phage display for binding to its peptide ligand. Binding properties of these variants defined a high-resolution map of mutational preference across the WW domain; each position had unique features that could not be captured by a few representative mutations. Our approach could be applied to many in vitro or in vivo protein assays, providing a general means for understanding how protein function relates to sequence.

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Más notícias para Darwin:

This can't be good news for Darwin: "...each position had unique features that could not be captured by a few representative mutations." If, indeed, each fitness peak lies separated by more than a few specific mutations, it remains difficult to envision how the Darwinian mechanism might facilitate the transition from one peak to another within any reasonable time frame.

Jonathan McLatchie