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PMID: 10430866 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Domain structure and lipid interaction of recombinant yeast Tim44.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 96 ·No. 16 ·1999-08-03 ·Pages 8890-4

Weiss C, Oppliger W, Vergères G, Demel R, Jenö P, Horst M, de Kruijff B, Schatz G, Azem A

Abstract

Tim44 is an essential component of the machinery that mediates the translocation of nuclear-encoded proteins across the mitochondrial inner membrane. It functions as a membrane anchor for the ATP-driven protein import motor whose other subunits are the mitochondrial 70-kDa heat-shock protein (mhsp70) and its nucleotide exchange factor, mGrpE. To understand how this motor is anchored to the inner membrane, we have overexpressed Tim44 in Escherichia coli and studied the properties of the pure protein and its interaction with model lipid membranes. Limited proteolysis and analytical ultracentrifugation indicate that Tim44 is an elongated monomer with a stably folded C-terminal domain. The protein binds strongly to liposomes composed of phosphatidylcholine and cardiolipin but only weakly to liposomes containing phosphatidylcholine alone. Studies with phospholipid monolayers suggest that Tim44 binds to phospholipids of the mitochondrial inner membrane both by electrostatic interactions and by penetrating the polar head group region.

MeSH Terms
Cardiolipins/metabolism Carrier Proteins/chemistry,genetics,metabolism Cloning, Molecular Escherichia coli Kinetics Liposomes/metabolism Macromolecular Substances Membrane Lipids/chemistry,metabolism Membrane Proteins/chemistry,genetics,metabolism Mitochondrial Membrane Transport Proteins Mitochondrial Precursor Protein Import Complex Proteins Open Reading Frames Phosphatidylcholines/metabolism Recombinant Proteins/chemistry,isolation & purification,metabolism Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins Sucrose/metabolism
Chemicals
Cardiolipins Carrier Proteins Liposomes Macromolecular Substances Membrane Lipids Membrane Proteins Mitochondrial Membrane Transport Proteins Mitochondrial Precursor Protein Import Complex Proteins Phosphatidylcholines Recombinant Proteins Saccharomyces cerevisiae Proteins TIM44 protein, S cerevisiae Sucrose 1,2-oleoylphosphatidylcholine
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Weiss C
Department of Biochemistry, Tel-Aviv University, Tel-Aviv 69978, Israel.
Oppliger W
Vergères G
Demel R
Jenö P
Horst M
de Kruijff B
Schatz G
Azem A
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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.96.16.8890
Published
1999-08-03
Pages
8890-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC17703
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.96.16.8890 PMC Full Text View journal details
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