Multiprotein complex production in E. coli: the SecYEG-SecDFYajC-YidC Holotranslocon

Imre Berger; Quiyang  Jiang; Ryan Schulze; Ian Collinson; Christiane Schaffitzel

doi:10.1007/978-1-4939-6887-9_18

Multiprotein complex production in E. coli: the SecYEG-SecDFYajC-YidC Holotranslocon

Imre Berger, Quiyang Jiang, Ryan Schulze, Ian Collinson, Christiane Schaffitzel

Research output: Chapter in Book/Report/Conference proceeding › Chapter in a book

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Abstract

A modular approach for balanced overexpression of recombinant multiprotein complexes in E. coli is described, with the prokaryotic protein secretase/insertase complex, the SecYEG-SecDFYajC-YidC holotranslocon (HTL), used as an example. This procedure has been implemented here in the ACEMBL system. The protocol details the design principles of the monocistronic or polycistronic DNA constructs, the expression and purification of functional HTL and its association with translating ribosome nascent chain (RNC) complexes into a RNC-HTL supercomplex.

Original language	English
Title of host publication	Heterologous Gene Expression in E.coli
Subtitle of host publication	Methods and Protocols
Editors	Nichola A Burgess-Brown
Publisher	Springer
Pages	279-290
Number of pages	12
ISBN (Electronic)	9781493968879
ISBN (Print)	9781493968855
DOIs	https://doi.org/10.1007/978-1-4939-6887-9_18
Publication status	Published - 2017

Publication series

Name	Methods in Molecular Biology
Publisher	Springer New York
Volume	1586
ISSN (Print)	1064-3745

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10.1007/978-1-4939-6887-9_18

Full-text PDF (accepted author manuscript)
This is the author accepted manuscript (AAM). The final published version (version of record) is available online via Springer at https://link.springer.com/protocol/10.1007%2F978-1-4939-6887-9_18. Please refer to any applicable terms of use of the publisher.
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Membrane protein insertion and quality control by the bacterial holo-translocon and FtsH chaperone/protease complex
Berger-Schaffitzel, C. H. (Principal Investigator)
9/01/17 → 8/07/20
Project: Research
Understanding the Mechanism of Membrane Protein Insertion
Collinson, I. R. (Principal Investigator)
1/10/14 → 28/02/18
Project: Research
Ensemble and Single Molecule Analysis of Protein Translocation
Collinson, I. R. (Principal Investigator)
1/01/12 → 1/01/16
Project: Research

Cite this

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abstract = "A modular approach for balanced overexpression of recombinant multiprotein complexes in E. coli is described, with the prokaryotic protein secretase/insertase complex, the SecYEG-SecDFYajC-YidC holotranslocon (HTL), used as an example. This procedure has been implemented here in the ACEMBL system. The protocol details the design principles of the monocistronic or polycistronic DNA constructs, the expression and purification of functional HTL and its association with translating ribosome nascent chain (RNC) complexes into a RNC-HTL supercomplex.",

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Multiprotein complex production in E. coli: the SecYEG-SecDFYajC-YidC Holotranslocon. / Berger, Imre; Jiang, Quiyang ; Schulze, Ryan et al.
Heterologous Gene Expression in E.coli: Methods and Protocols. ed. / Nichola A Burgess-Brown. Springer, 2017. p. 279-290 (Methods in Molecular Biology; Vol. 1586).

Research output: Chapter in Book/Report/Conference proceeding › Chapter in a book

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T1 - Multiprotein complex production in E. coli

T2 - the SecYEG-SecDFYajC-YidC Holotranslocon

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AU - Jiang, Quiyang

AU - Schulze, Ryan

AU - Collinson, Ian

AU - Schaffitzel, Christiane

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AB - A modular approach for balanced overexpression of recombinant multiprotein complexes in E. coli is described, with the prokaryotic protein secretase/insertase complex, the SecYEG-SecDFYajC-YidC holotranslocon (HTL), used as an example. This procedure has been implemented here in the ACEMBL system. The protocol details the design principles of the monocistronic or polycistronic DNA constructs, the expression and purification of functional HTL and its association with translating ribosome nascent chain (RNC) complexes into a RNC-HTL supercomplex.

U2 - 10.1007/978-1-4939-6887-9_18

DO - 10.1007/978-1-4939-6887-9_18

M3 - Chapter in a book

C2 - 28470612

SN - 9781493968855

T3 - Methods in Molecular Biology

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BT - Heterologous Gene Expression in E.coli

A2 - Burgess-Brown, Nichola A

PB - Springer

ER -

Multiprotein complex production in E. coli: the SecYEG-SecDFYajC-YidC Holotranslocon

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Projects

Membrane protein insertion and quality control by the bacterial holo-translocon and FtsH chaperone/protease complex

Understanding the Mechanism of Membrane Protein Insertion

Ensemble and Single Molecule Analysis of Protein Translocation

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