Production of Multi-subunit Membrane Protein Complexes

Burak V Kabasakal, Qiyang Jiang, Christiane H Berger-Schaffitzel

Research output: Chapter in Book/Report/Conference proceedingChapter in a book

Abstract

Membrane proteins constitute an important class of proteins for medical, pharmaceutical, and biotechnological reasons. Understanding the structure and function of membrane proteins and their complexes is of key importance, but the progress in this area is slow because of the difficulties to produce them in sufficient quality and quantity. Overexpression of membrane proteins is often restricted by the limited capability of translocation systems to integrate proteins into the membrane and to fold them properly. Purification of membrane proteins requires their isolation from the membrane, which is a further challenge. The choice of expression system, detergents, and purification tags is therefore an important decision. Here, we present a protocol for expression in bacteria and isolation of a seven-subunit membrane protein complex, the bacterial holo-translocon, which can serve as a starting point for the production of other membrane protein complexes for structural and functional studies.
Original languageEnglish
Title of host publicationMultiprotein Complexes
Subtitle of host publicationMethods and Protocols
EditorsArnaud Poterszman
PublisherSpringer US
Chapter1
Pages3-16
Volume2247
Edition1
ISBN (Electronic)978-1-0716-1126-5
ISBN (Print)978-1-0716-1125-8
DOIs
Publication statusPublished - 11 Dec 2020

Keywords

  • Membrane protein complex
  • Expression in bacteria
  • Endogenous host
  • Complex purification
  • Translocon

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