Purification and Functional Reconstitution of the Bacterial Protein Translocation Pore, the SecYEG Complex

I. Kusters; G. van den Bogaart; J.  de Wit; V. Krasnikov; B. Poolman; A. Driessen

doi:10.1007/978-1-60327-412-8_8

Purification and Functional Reconstitution of the Bacterial Protein Translocation Pore, the SecYEG Complex

I. Kusters, G. van den Bogaart, J. de Wit, V. Krasnikov, B. Poolman, A. Driessen

Research output: Chapter in Book/Report/Conference proceeding › Chapter › Academic › peer-review

9 Citations (Scopus)

Abstract

In bacteria, proteins are secreted across the cytoplasmic membrane by a protein complex termed translocase. The ability to study the activity of the translocase in vitro using purified proteins has been instrumental for our understanding of the mechanisms underlying this process. Here, we describe the protocols for the purification and reconstitution of the SecYEG complex in an active state into liposomes. In addition, fluorescence based in vitro assays are described that allow monitoring translocation activity discontinuously and in real time.

Original language	English
Title of host publication	Protein Secretion
Subtitle of host publication	Methods and Protocols
Editors	Anastassios Economou
Publisher	Humana Press
Chapter	8
Pages	131-143
Number of pages	13
Volume	619
ISBN (Electronic)	978-1-60327-412-8
ISBN (Print)	978-1-60327-167-7, 978-1-4939-5707-1
DOIs	https://doi.org/10.1007/978-1-60327-412-8_8
Publication status	Published - 25-Jan-2010

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Purification and Functional Reconstitution of the Bacterial Protein Translocation Pore, the SecYEG Complex
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Purification and Functional Reconstitution of the Bacterial Protein Translocation Pore, the SecYEG Complex. / Kusters, I.; Bogaart, G. van den ; de Wit, J. et al.
Protein Secretion: Methods and Protocols. ed. / Anastassios Economou. Vol. 619 Humana Press, 2010. p. 131-143.

Research output: Chapter in Book/Report/Conference proceeding › Chapter › Academic › peer-review

TY - CHAP

T1 - Purification and Functional Reconstitution of the Bacterial Protein Translocation Pore, the SecYEG Complex

AU - Kusters, I.

AU - Bogaart, G. van den

AU - de Wit, J.

AU - Krasnikov, V.

AU - Poolman, B.

AU - Driessen, A.

N1 - Relation: http://www.springerlink.com/ Rights: Springer

PY - 2010/1/25

Y1 - 2010/1/25

N2 - In bacteria, proteins are secreted across the cytoplasmic membrane by a protein complex termed translocase. The ability to study the activity of the translocase in vitro using purified proteins has been instrumental for our understanding of the mechanisms underlying this process. Here, we describe the protocols for the purification and reconstitution of the SecYEG complex in an active state into liposomes. In addition, fluorescence based in vitro assays are described that allow monitoring translocation activity discontinuously and in real time.

AB - In bacteria, proteins are secreted across the cytoplasmic membrane by a protein complex termed translocase. The ability to study the activity of the translocase in vitro using purified proteins has been instrumental for our understanding of the mechanisms underlying this process. Here, we describe the protocols for the purification and reconstitution of the SecYEG complex in an active state into liposomes. In addition, fluorescence based in vitro assays are described that allow monitoring translocation activity discontinuously and in real time.

U2 - 10.1007/978-1-60327-412-8_8

DO - 10.1007/978-1-60327-412-8_8

M3 - Chapter

SN - 978-1-60327-167-7

SN - 978-1-4939-5707-1

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BT - Protein Secretion

A2 - Economou, Anastassios

PB - Humana Press

ER -

Purification and Functional Reconstitution of the Bacterial Protein Translocation Pore, the SecYEG Complex

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