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PMID: 2649482 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Temperature-dependent insertion of prolipoprotein into Escherichia coli membrane vesicles and requirements for ATP, soluble factors, and functional SecY protein for the overall translocation process.

Journal of bacteriology ·Vol. 171 ·No. 4 ·1989-04-00 ·Pages 1987-97

Tian G, Wu HC, Ray PH, Tai PC

Abstract

The requirements for the translocation of prolipoprotein into membrane vesicles were examined in an in vitro system. As measured by the eventual modification and processing of the prolipoprotein to form mature lipoprotein, the overall translocation process was found to require ATP hydrolysis, the presence of some heat-labile soluble cytoplasmic translocation factors, and the function of a cytoplasmic membrane protein, SecY/PrlA. However, the initial step of complete insertion of prolipoprotein into the membrane vesicles occurred without apparent requirements of a nucleotide, cytoplasmic translocation factors, or a functional SecY/PrlA membrane protein. Immunopurified prolipoprotein spontaneously inserted into membrane vesicles at elevated temperatures and required ATP and cytoplasmic translocation factors to form mature lipoprotein. The prolipoprotein inserted most efficiently into liposomes made of negatively charged phospholipids, indicating the importance of phospholipids in protein translocation. These results suggest that ATP hydrolysis and the actions of both cytoplasmic translocation factors and a functional SecY/PrlA membrane protein occur at a step(s) after the insertion of the precursors into membrane vesicles. The initial step of spontaneous insertion of prolipoprotein into membranes is in good agreement with membrane trigger hypothesis proposed by W. Wickner (Annu. Rev. Biochem. 48:23-45, 1979) and the helical hairpin hypothesis proposed by D. M. Engleman and T. A. Steitz (Cell 23:411-422, 1981).

MeSH Terms
Adenosine Triphosphate/metabolism Bacterial Outer Membrane Proteins/metabolism Bacterial Proteins/metabolism Biological Transport Cell Membrane/metabolism Cytoplasm/physiology Escherichia coli/metabolism Escherichia coli Proteins Lipoproteins/metabolism Liposomes Molecular Weight Protein Precursors/metabolism Protein Processing, Post-Translational Temperature
Chemicals
Bacterial Outer Membrane Proteins Bacterial Proteins Escherichia coli Proteins Lipoproteins Liposomes Lpp protein, E coli Protein Precursors Adenosine Triphosphate
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Tian G
Department of Metabolic Regulation, Boston Biomedical Research Institute, Massachusetts 02114.
Wu H C
Ray P H
Tai P C
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71 references, click to expand
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.171.4.1987-1997.1989
Published
1989-04-00
Pages
1987-97
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC209849
Subset
IM
Grants
DRS NIH HHS · BRSG 05511 · United States
NIGMS NIH HHS · GM28811 · United States
NIGMS NIH HHS · GM34538 · United States
External Links
PubMed source DOI: 10.1128/jb.171.4.1987-1997.1989 PMC Full Text View journal details
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