PUBLICATION
Adaptation of a Genetic Screen Reveals an Inhibitor for Mitochondrial Protein Import Component Tim44
- Authors
- Miyata, N., Tang, Z., Conti, M.A., Johnson, M.E., Douglas, C.J., Hasson, S.A., Damoiseaux, R., Chang, C.A., Koehler, C.M.
- ID
- ZDB-PUB-190321-19
- Date
- 2017
- Source
- The Journal of biological chemistry 292: 5429-5442 (Journal)
- Registered Authors
- Koehler, Carla
- Keywords
- chemical biology, mitochondria, mitochondrial transport, protein translocation, small molecule
- MeSH Terms
-
- Animals
- Binding Sites
- Genetic Testing
- HeLa Cells
- Humans
- Mitochondrial Membrane Transport Proteins/antagonists & inhibitors*
- Mitochondrial Membrane Transport Proteins/chemistry
- Mitochondrial Membrane Transport Proteins/metabolism
- Models, Molecular
- Molecular Docking Simulation
- Molecular Dynamics Simulation
- Neurospora crassa
- Protein Transport/drug effects
- Saccharomyces cerevisiae/drug effects*
- Saccharomyces cerevisiae Proteins/antagonists & inhibitors*
- Saccharomyces cerevisiae Proteins/chemistry
- Saccharomyces cerevisiae Proteins/metabolism
- Zebrafish
- PubMed
- 28167535 Full text @ J. Biol. Chem.
Citation
Miyata, N., Tang, Z., Conti, M.A., Johnson, M.E., Douglas, C.J., Hasson, S.A., Damoiseaux, R., Chang, C.A., Koehler, C.M. (2017) Adaptation of a Genetic Screen Reveals an Inhibitor for Mitochondrial Protein Import Component Tim44. The Journal of biological chemistry. 292:5429-5442.
Abstract
Diverse protein import pathways into mitochondria use translocons on the outer membrane (TOM) and inner membrane (TIM). We adapted a genetic screen, based on Ura3 mistargeting from mitochondria to the cytosol, to identify small molecules that attenuated protein import. Small molecule mitochondrial import blockers of the Carla Koehler laboratory (MB)-10 inhibited import of substrates that require the TIM23 translocon. Mutational analysis coupled with molecular docking and molecular dynamics modeling revealed that MB-10 binds to a specific pocket in the C-terminal domain of Tim44 of the protein-associated motor (PAM) complex. This region was proposed to anchor Tim44 to the membrane, but biochemical studies with MB-10 show that this region is required for binding to the translocating precursor and binding to mtHsp70 in low ATP conditions. This study also supports a direct role for the PAM complex in the import of substrates that are laterally sorted to the inner membrane, as well as the mitochondrial matrix. Thus, MB-10 is the first small molecule modulator to attenuate PAM complex activity, likely through binding to the C-terminal region of Tim44.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping