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Fine Sampling of Sequence Space for Membrane Protein Structural Biology

J Mol Biol. 2021-05; 
Michael Loukeris, Zahra Assur Sanghai, Jeremie Vendome, Wayne A Hendrickson, Brian Kloss, Filippo Mancia
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Gene Synthesis Target sequences were selected from the NYCOMPS genome library using a bioinformatics approach,11, 12 or synthesized commercially (GenScript) Get A Quote

摘要

We describe an enhancement of traditional genomics-based approaches to improve the success of structure determination of membrane proteins. Following a broad screen of sequence space to identify initial expression-positive targets, we employ a second step to select orthologs with closely related sequences to these hits. We demonstrate that a greater percentage of these latter targets express well and are stable in detergent, increasing the likelihood of identifying candidates that will ultimately yield structural information.

关键词

high throughput biology, integral membrane proteins, protein expression, protein purification, structural biology, structural genomics