High-throughput expression and purification of membrane proteins

Filippo Mancia, James Love

Research output: Contribution to journalArticle

26 Scopus citations

Abstract

High-throughput (HT) methodologies have had a tremendous impact on structural biology of soluble proteins. High-resolution structure determination relies on the ability of the macromolecule to form ordered crystals that diffract X-rays. While crystallization remains somewhat empirical, for a given protein, success is proportional to the number of conditions screened and to the number of variants trialed. HT techniques have greatly increased the number of targets that can be trialed and the rate at which these can be produced. In terms of number of structures solved, membrane proteins appear to be lagging many years behind their soluble counterparts. Likewise, HT methodologies for production and characterization of these hydrophobic macromolecules are only now emerging. Presented here is an HT platform designed exclusively for membrane proteins that has processed over 5000 targets.

Original languageEnglish (US)
Pages (from-to)85-93
Number of pages9
JournalJournal of Structural Biology
Volume172
Issue number1
DOIs
Publication statusPublished - Oct 1 2010

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Keywords

  • Detergent
  • Expression
  • High-throughput
  • Membrane proteins
  • Structural genomics
  • Structure

ASJC Scopus subject areas

  • Structural Biology

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