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PMID: 2554321 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Specific recognition of the leader region of precursor proteins is required for the activation of translocation ATPase of Escherichia coli.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 86 ·No. 22 ·1989-11-00 ·Pages 8630-4

Cunningham K, Wickner W

Abstract

The ATP-hydrolytic activity of SecA protein is stimulated up to 100-fold by the translocation-competent precursor to outer membrane protein A (pro-OmpA) in conjunction with inner-membrane vesicles bearing active SecY [Lill, R., Cunningham, K., Brundage, L., Ito, K., Oliver, D. & Wickner, W. (1989) EMBO J. 8, 961-966]. This reaction is saturable, with Michaelis-Menten kinetics for an enzyme with two substrates, ATP and pro-OmpA, and is defined as translocation ATPase. Another precursor protein, pre-PhoE, is also a substrate for this translocation ATPase. Neither OmpA nor its synthetic leader peptide are effective substrates for translocation ATPase, suggesting that both domains of the complete precursor are necessary for the reaction. The leader peptide is a potent inhibitor and apparently competes with pro-OmpA for necessary binding sites on translocation ATPase. After a brief preincubation, the activity of translocation ATPase becomes resistant to inhibition by leader peptide, suggesting that the leader peptide is recognized at an early step in the protein translocation pathway. Our enzymological studies show that translocation ATPase recognizes and functionally binds the leader region of precursor proteins.

MeSH Terms
Amino Acid Sequence Bacterial Outer Membrane Proteins/metabolism Bacterial Proteins/metabolism Cell Membrane/enzymology Enzyme Activation Escherichia coli/enzymology Kinetics Molecular Sequence Data Oligopeptides Protein Sorting Signals/metabolism Proton-Translocating ATPases/metabolism
Chemicals
Bacterial Outer Membrane Proteins Bacterial Proteins Oligopeptides Protein Sorting Signals Proton-Translocating ATPases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Cunningham K
Molecular Biology Institute, University of California, Los Angeles 90024-1570.
Wickner W
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34 references, click to expand
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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.86.22.8630
Published
1989-11-00
Pages
8630-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC298341
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.86.22.8630 PMC Full Text View journal details
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