Motif-directed Redesign of Enzyme Specificity

Borgo B and Havranek JJ, 23(3):312-20, Protein Sci, 2014

A detailed computational sequence analysis examining specific residues and motifs of a methoionine aminopeptidase (eMAP) from E. coli was performed in attempts to better understand the specificity and binding activity of this enzyme. The goal was to switch the specificity from N-terminal methionine to N-terminal leucine using predicted motif-based redesign. Binding kinetics on the native and engineered proteins were performed on the BLItz system using Amine-Reactive biosensors. This motif-based computational design combined with a focused functional study provides protein engineers with new tools to more rapidly develop and test enzyme model systems.

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