Transforming a (beta/alpha)8--barrel enzyme into a split-protein sensor through directed evolution.

Petra Tafelmeyer; Nils Johnsson; Kai Johnsson

doi:10.1016/j.chembiol.2004.02.026

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Transforming a (beta/alpha)8--barrel enzyme into a split-protein sensor through directed evolution.

Petra Tafelmeyer·Nils Johnsson·Kai Johnsson

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Abstract

Split-protein sensors have become an important tool for the analysis of protein-protein interactions in living cells. We present here a combinatorial method for the generation of new split-protein sensors and demonstrate its application toward the (beta/alpha)(8)-barrel enzyme N-(5'-phosphoribosyl)-anthranilate isomerase Trp1p from Saccharomyces cerevisiae. The generated split-Trp protein sensors allow for the detection of protein-protein interactions in the cytosol as well as the membrane by enabling trp1 cells to grow on medium lacking tryptophan. This powerful selection complements the repertoire of the currently used split-protein sensors and provides a new tool for high-throughput interaction screening.

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Aldose-Ketose Isomerases Amino Acid Sequence Binding Sites Biosensing Techniques Combinatorial Chemistry Techniques Directed Molecular Evolution Evolution, Molecular Membrane Proteins Molecular Sequence Data Peptide Fragments Protein Interaction Mapping Protein Structure, Secondary Proteins Recombinant Fusion Proteins Saccharomyces cerevisiae Sequence Deletion Tryptophan Journal Article Research Support, Non-U.S. Gov't

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Petra Tafelmeyer, Nils Johnsson, Kai Johnsson. 2004. Transforming a (beta/alpha)8--barrel enzyme into a split-protein sensor through directed evolution.. https://doi.org/10.1016/j.chembiol.2004.02.026

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