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PMID: 1830665 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Reconstitution of a protein translocation system containing purified SecY, SecE, and SecA from Escherichia coli.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 88 ·No. 15 ·1991-08-01 ·Pages 6545-9

Akimaru J, Matsuyama S, Tokuda H, Mizushima S

Abstract

Reconstitution of the translocation machinery for secretory proteins from purified constituents was performed. SecY was solubilized from SecY/SecE-overproducing Escherichia coli cells and purified by chromatography on ion-exchange and size-exclusion columns. Proteoliposomes active in protein translocation were reconstituted from the purified preparations of SecY and SecE. The reconstituted translocation activity was SecA- and ATP-dependent. Although the purified preparations of SecY and SecE were still contaminated with minute amounts of other proteins, the elution profiles of SecY and SecE on column chromatographies coincided with the elution profiles of reconstituted translocation activity, indicating that SecY and SecE are the indispensable components in these preparations. We conclude that SecY, SecE, and SecA are essential components of the protein secretion machinery and that translocation activity can be reconstituted from only these three proteins and phospholipids.

MeSH Terms

Adenosine Triphosphatases/isolation & purification,metabolism Bacterial Outer Membrane Proteins/genetics,metabolism Bacterial Proteins/isolation & purification,metabolism Cell Membrane/metabolism Chromatography, Gel Chromatography, Ion Exchange Escherichia coli/genetics,metabolism Escherichia coli Proteins Genes, Bacterial Kinetics Liposomes Membrane Transport Proteins Molecular Weight Phospholipids/metabolism Protein Biosynthesis Proteolipids/metabolism SEC Translocation Channels SecA Proteins Transcription, Genetic

Chemicals

Bacterial Outer Membrane Proteins Bacterial Proteins Escherichia coli Proteins Liposomes Membrane Transport Proteins Phospholipids Proteolipids SEC Translocation Channels SecE protein, E coli SecY protein, E coli proteoliposomes Adenosine Triphosphatases SecA Proteins

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

Akimaru J

Institute of Applied Microbiology, University of Tokyo, Japan.

Matsuyama S

Tokuda H

Mizushima S

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Article Info

Journal

Proceedings of the National Academy of Sciences of the United States of America

Abbr.

Proc Natl Acad Sci U S A

ISSN

0027-8424

DOI

10.1073/pnas.88.15.6545

Published

1991-08-01

Pages

6545-9

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC52123

Subset

External Links

PubMed source DOI: 10.1073/pnas.88.15.6545 PMC Full Text View journal details

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Reconstitution of a protein translocation system containing purified SecY, SecE, and SecA from Escherichia coli.

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