Adaptation of a genetic screen reveals an inhibitor for mitochondrial protein import component Tim44

Non Miyata, Zhiye Tang, Michael A. Conti, Meghan E. Johnson, Colin J. Douglas, Samuel A. Hasson, Robert Damoiseaux, Chia En A. Chang, Carla M. Koehler

研究成果: ジャーナルへの寄稿記事

2 引用 (Scopus)

抄録

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.

元の言語英語
ページ(範囲)5429-5442
ページ数14
ジャーナルJournal of Biological Chemistry
292
発行部数13
DOI
出版物ステータス出版済み - 3 31 2017

Fingerprint

Mitochondrial Proteins
Membranes
Mitochondria
Proteins
Molecules
Molecular Dynamics Simulation
Cytosol
Substrates
Motor Activity
Anchors
Adenosine Triphosphate
Modulators
Molecular dynamics

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

これを引用

Miyata, N., Tang, Z., Conti, M. A., Johnson, M. E., Douglas, C. J., Hasson, S. A., ... Koehler, C. M. (2017). Adaptation of a genetic screen reveals an inhibitor for mitochondrial protein import component Tim44. Journal of Biological Chemistry, 292(13), 5429-5442. https://doi.org/10.1074/jbc.M116.770131

Adaptation of a genetic screen reveals an inhibitor for mitochondrial protein import component Tim44. / Miyata, Non; Tang, Zhiye; Conti, Michael A.; Johnson, Meghan E.; Douglas, Colin J.; Hasson, Samuel A.; Damoiseaux, Robert; Chang, Chia En A.; Koehler, Carla M.

:: Journal of Biological Chemistry, 巻 292, 番号 13, 31.03.2017, p. 5429-5442.

研究成果: ジャーナルへの寄稿記事

Miyata, N, Tang, Z, Conti, MA, Johnson, ME, Douglas, CJ, Hasson, SA, Damoiseaux, R, Chang, CEA & Koehler, CM 2017, 'Adaptation of a genetic screen reveals an inhibitor for mitochondrial protein import component Tim44', Journal of Biological Chemistry, 巻. 292, 番号 13, pp. 5429-5442. https://doi.org/10.1074/jbc.M116.770131
Miyata, Non ; Tang, Zhiye ; Conti, Michael A. ; Johnson, Meghan E. ; Douglas, Colin J. ; Hasson, Samuel A. ; Damoiseaux, Robert ; Chang, Chia En A. ; Koehler, Carla M. / Adaptation of a genetic screen reveals an inhibitor for mitochondrial protein import component Tim44. :: Journal of Biological Chemistry. 2017 ; 巻 292, 番号 13. pp. 5429-5442.
@article{61d230cfe1954905850cf8d2914a1627,
title = "Adaptation of a genetic screen reveals an inhibitor for mitochondrial protein import component Tim44",
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.",
author = "Non Miyata and Zhiye Tang and Conti, {Michael A.} and Johnson, {Meghan E.} and Douglas, {Colin J.} and Hasson, {Samuel A.} and Robert Damoiseaux and Chang, {Chia En A.} and Koehler, {Carla M.}",
year = "2017",
month = "3",
day = "31",
doi = "10.1074/jbc.M116.770131",
language = "English",
volume = "292",
pages = "5429--5442",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "13",

}

TY - JOUR

T1 - Adaptation of a genetic screen reveals an inhibitor for mitochondrial protein import component Tim44

AU - Miyata, Non

AU - Tang, Zhiye

AU - Conti, Michael A.

AU - Johnson, Meghan E.

AU - Douglas, Colin J.

AU - Hasson, Samuel A.

AU - Damoiseaux, Robert

AU - Chang, Chia En A.

AU - Koehler, Carla M.

PY - 2017/3/31

Y1 - 2017/3/31

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85016608155&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85016608155&partnerID=8YFLogxK

U2 - 10.1074/jbc.M116.770131

DO - 10.1074/jbc.M116.770131

M3 - Article

VL - 292

SP - 5429

EP - 5442

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 13

ER -