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Methods Mol Biol. 2014;1140:201-9. doi: 10.1007/978-1-4939-0354-2_16.

Salvage or recovery of failed targets by mutagenesis to reduce surface entropy.

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1
UCLA-DOE Institute for Genomics and Proteomics, Howard Hughes Medical Institute, University of California, Los Angeles, CA, USA.

Abstract

The success of macromolecular crystallization depends on the protein's ability to form specific, cohesive intermolecular interactions that serve as crystal contacts. In the cases where the protein lacks surface patches conducive to such interactions, crystallization may not occur. However, it is possible to enhance the likelihood of crystallization by engineering such patches through site-directed mutagenesis, targeting specifically residues with high side chain entropy and replacing them with small amino acids (i.e., surface entropy reduction, SER). This method has proven successful in hundreds of crystallographic analyses of proteins otherwise recalcitrant to crystallization. Three representative cases of the application of the SER strategy, assisted by the automated prediction of the mutation sites using the SER prediction (SERp) server are described.

PMID:
24590720
DOI:
10.1007/978-1-4939-0354-2_16
[Indexed for MEDLINE]
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