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Please use this identifier to cite or link to this item: http://hdl.handle.net/10481/28541

Title: Probing the Mutational Interplay between Primary and Promiscuous Protein Functions: A Computational-Experimental Approach
Authors: García Seisdedos, Héctor
Ibarra Molero, Beatriz
Sánchez Ruiz, José Manuel
Issue Date: 2012
Abstract: Protein promiscuity is of considerable interest due its role in adaptive metabolic plasticity, its fundamental connection with molecular evolution and also because of its biotechnological applications. Current views on the relation between primary and promiscuous protein activities stem largely from laboratory evolution experiments aimed at increasing promiscuous activity levels. Here, on the other hand, we attempt to assess the main features of the simultaneous modulation of the primary and promiscuous functions during the course of natural evolution. The computational/experimental approach we propose for this task involves the following steps: a function-targeted, statistical coupling analysis of evolutionary data is used to determine a set of positions likely linked to the recruitment of a promiscuous activity for a new function; a combinatorial library of mutations on this set of positions is prepared and screened for both, the primary and the promiscuous activities; a partial-least-squares reconstruction of the full combinatorial space is carried out; finally, an approximation to the Pareto set of variants with optimal primary/promiscuous activities is derived. Application of the approach to the emergence of folding catalysis in thioredoxin scaffolds reveals an unanticipated scenario: diverse patterns of primary/promiscuous activity modulation are possible, including a moderate (but likely significant in a biological context) simultaneous enhancement of both activities. We show that this scenario can be most simply explained on the basis of the conformational diversity hypothesis, although alternative interpretations cannot be ruled out. Overall, the results reported may help clarify the mechanisms of the evolution of new functions. From a different viewpoint, the partial-least-squares-reconstruction/Par​eto-set-predictionapproach we have introduced provides the computational basis for an efficient directed-evolution protocol aimed at the simultaneous enhancement of several protein features and should therefore open new possibilities in the engineering of multi-functional enzymes.
Sponsorship: This work was supported by FEDER Funds and Grants BIO2009-09562 and CSD2009-00088 from the Spanish Ministry of Science and Innovation.
Publisher: Public Library of Science (PLOS)
Keywords: Sequence analysis
Library screening
Mutation
Protein folding
Enzymes
Biocatalysis
Sequence alignment
Molecular evolution
URI: http://hdl.handle.net/10481/28541
ISSN: 1553-734X
1553-7358
Rights : Creative Commons Attribution-NonCommercial-NoDerivs 3.0 License
Citation: García Seisdedos, H.; Ibarra Molero, B.; Sánchez Ruiz, J.M. Probing the Mutational Interplay between Primary and Promiscuous Protein Functions: A Computational-Experimental Approach. Plos Computational Biology, 8(6): e1002558 (2012). [http://hdl.handle.net/10481/28541]
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