Mechanism of human PINK1 activation at the TOM complex in a reconstituted system

Olawale G. Raimi; Hina Ojha; Kenneth Ehses; Verena Dederer; Sven M. Lange; Cristian Polo Rivera; Tom D. Deegan; Yinchen Chen; Melanie Wightman; Rachel Toth; Karim P. M. Labib; Sebastian Mathea; Neil Ranson; Rubén Fernández-Busnadiego; Miratul M. K. Muqit

doi:10.1126/sciadv.adn7191

Mechanism of human PINK1 activation at the TOM complex in a reconstituted system

Olawale G. Raimi, Hina Ojha, Kenneth Ehses, Verena Dederer, Sven M. Lange, Cristian Polo Rivera, Tom D. Deegan, Yinchen Chen, Melanie Wightman, Rachel Toth, Karim P. M. Labib, Sebastian Mathea, Neil Ranson, Rubén Fernández-Busnadiego, Miratul M. K. Muqit (Lead / Corresponding author)

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Abstract

Loss-of-function mutations in PTEN-induced kinase 1 (PINK1) are a frequent cause of early-onset Parkinson's disease (PD). Stabilization of PINK1 at the translocase of outer membrane (TOM) complex of damaged mitochondria is critical for its activation. The mechanism of how PINK1 is activated in the TOM complex is unclear. Here, we report that co-expression of human PINK1 and all seven TOM subunits in Saccharomyces cerevisiae is sufficient for PINK1 activation. We use this reconstitution system to systematically assess the role of each TOM subunit toward PINK1 activation. We unambiguously demonstrate that the TOM20 and TOM70 receptor subunits are required for optimal PINK1 activation and map their sites of interaction with PINK1 using AlphaFold structural modeling and mutagenesis. We also demonstrate an essential role of the pore-containing subunit TOM40 and its structurally associated subunits TOM7 and TOM22 for PINK1 activation. These findings will aid in the development of small-molecule activators of PINK1 as a therapeutic strategy for PD.

Original language	English
Article number	eadn7191
Number of pages	16
Journal	Science Advances
Volume	10
Issue number	23
Early online date	7 Jun 2024
DOIs	https://doi.org/10.1126/sciadv.adn7191
Publication status	Published - Jun 2024

Keywords

Protein Kinases/metabolism
Humans
Mitochondrial Precursor Protein Import Complex Proteins/metabolism
Mitochondrial Membrane Transport Proteins/metabolism
Saccharomyces cerevisiae/metabolism
Saccharomyces cerevisiae Proteins/metabolism
Mitochondria/metabolism
Protein Binding
Enzyme Activation
Models, Molecular
Protein Subunits/metabolism

ASJC Scopus subject areas

General

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1126/sciadv.adn7191Licence: CC BY-NC

Final published versionFinal published version, 1.78 MBLicence: CC BY-NC

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Raimi, O. G., Ojha, H., Ehses, K., Dederer, V., Lange, S. M., Rivera, C. P., Deegan, T. D., Chen, Y., Wightman, M., Toth, R., Labib, K. P. M., Mathea, S., Ranson, N., Fernández-Busnadiego, R., & Muqit, M. M. K. (2024). Mechanism of human PINK1 activation at the TOM complex in a reconstituted system. Science Advances, 10(23), Article eadn7191. https://doi.org/10.1126/sciadv.adn7191

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title = "Mechanism of human PINK1 activation at the TOM complex in a reconstituted system",

abstract = "Loss-of-function mutations in PTEN-induced kinase 1 (PINK1) are a frequent cause of early-onset Parkinson's disease (PD). Stabilization of PINK1 at the translocase of outer membrane (TOM) complex of damaged mitochondria is critical for its activation. The mechanism of how PINK1 is activated in the TOM complex is unclear. Here, we report that co-expression of human PINK1 and all seven TOM subunits in Saccharomyces cerevisiae is sufficient for PINK1 activation. We use this reconstitution system to systematically assess the role of each TOM subunit toward PINK1 activation. We unambiguously demonstrate that the TOM20 and TOM70 receptor subunits are required for optimal PINK1 activation and map their sites of interaction with PINK1 using AlphaFold structural modeling and mutagenesis. We also demonstrate an essential role of the pore-containing subunit TOM40 and its structurally associated subunits TOM7 and TOM22 for PINK1 activation. These findings will aid in the development of small-molecule activators of PINK1 as a therapeutic strategy for PD.",

keywords = "Protein Kinases/metabolism, Humans, Mitochondrial Precursor Protein Import Complex Proteins/metabolism, Mitochondrial Membrane Transport Proteins/metabolism, Saccharomyces cerevisiae/metabolism, Saccharomyces cerevisiae Proteins/metabolism, Mitochondria/metabolism, Protein Binding, Enzyme Activation, Models, Molecular, Protein Subunits/metabolism",

author = "Raimi, {Olawale G.} and Hina Ojha and Kenneth Ehses and Verena Dederer and Lange, {Sven M.} and Rivera, {Cristian Polo} and Deegan, {Tom D.} and Yinchen Chen and Melanie Wightman and Rachel Toth and Labib, {Karim P. M.} and Sebastian Mathea and Neil Ranson and Rub{\'e}n Fern{\'a}ndez-Busnadiego and Muqit, {Miratul M. K.}",

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year = "2024",

month = jun,

doi = "10.1126/sciadv.adn7191",

language = "English",

volume = "10",

journal = "Science Advances",

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AU - Raimi, Olawale G.

AU - Ojha, Hina

AU - Ehses, Kenneth

AU - Dederer, Verena

AU - Lange, Sven M.

AU - Rivera, Cristian Polo

AU - Deegan, Tom D.

AU - Chen, Yinchen

AU - Wightman, Melanie

AU - Toth, Rachel

AU - Labib, Karim P. M.

AU - Mathea, Sebastian

AU - Ranson, Neil

AU - Fernández-Busnadiego, Rubén

AU - Muqit, Miratul M. K.

PY - 2024/6

Y1 - 2024/6

N2 - Loss-of-function mutations in PTEN-induced kinase 1 (PINK1) are a frequent cause of early-onset Parkinson's disease (PD). Stabilization of PINK1 at the translocase of outer membrane (TOM) complex of damaged mitochondria is critical for its activation. The mechanism of how PINK1 is activated in the TOM complex is unclear. Here, we report that co-expression of human PINK1 and all seven TOM subunits in Saccharomyces cerevisiae is sufficient for PINK1 activation. We use this reconstitution system to systematically assess the role of each TOM subunit toward PINK1 activation. We unambiguously demonstrate that the TOM20 and TOM70 receptor subunits are required for optimal PINK1 activation and map their sites of interaction with PINK1 using AlphaFold structural modeling and mutagenesis. We also demonstrate an essential role of the pore-containing subunit TOM40 and its structurally associated subunits TOM7 and TOM22 for PINK1 activation. These findings will aid in the development of small-molecule activators of PINK1 as a therapeutic strategy for PD.

AB - Loss-of-function mutations in PTEN-induced kinase 1 (PINK1) are a frequent cause of early-onset Parkinson's disease (PD). Stabilization of PINK1 at the translocase of outer membrane (TOM) complex of damaged mitochondria is critical for its activation. The mechanism of how PINK1 is activated in the TOM complex is unclear. Here, we report that co-expression of human PINK1 and all seven TOM subunits in Saccharomyces cerevisiae is sufficient for PINK1 activation. We use this reconstitution system to systematically assess the role of each TOM subunit toward PINK1 activation. We unambiguously demonstrate that the TOM20 and TOM70 receptor subunits are required for optimal PINK1 activation and map their sites of interaction with PINK1 using AlphaFold structural modeling and mutagenesis. We also demonstrate an essential role of the pore-containing subunit TOM40 and its structurally associated subunits TOM7 and TOM22 for PINK1 activation. These findings will aid in the development of small-molecule activators of PINK1 as a therapeutic strategy for PD.

KW - Protein Kinases/metabolism

KW - Humans

KW - Mitochondrial Precursor Protein Import Complex Proteins/metabolism

KW - Mitochondrial Membrane Transport Proteins/metabolism

KW - Saccharomyces cerevisiae/metabolism

KW - Saccharomyces cerevisiae Proteins/metabolism

KW - Mitochondria/metabolism

KW - Protein Binding

KW - Enzyme Activation

KW - Models, Molecular

KW - Protein Subunits/metabolism

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U2 - 10.1126/sciadv.adn7191

DO - 10.1126/sciadv.adn7191

M3 - Article

C2 - 38848361

SN - 2375-2548

VL - 10

JO - Science Advances

JF - Science Advances

IS - 23

M1 - eadn7191

ER -

Mechanism of human PINK1 activation at the TOM complex in a reconstituted system

Abstract

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

Other files and links

Fingerprint

ASAP - Mapping the LRRK2 Signalling Pathway and its Interplay with other Parkinson's Disease Components

Development of better approaches to study PINK1 structure (joint application with University Medical Centre Goettingen and University of Frankfurt)

Novel Genome Integrity Pathways that Regulate DNA Replication Termination in Metazoa

Mechanism of human PINK1 activation at the TOM complex by reconstitution

MRC PPU DNA Sequencing and Services

MRC PPU Reagents and Services

Cite this

Mechanism of human PINK1 activation at the TOM complex in a reconstituted system

Abstract

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

Other files and links

Fingerprint

Projects

ASAP - Mapping the LRRK2 Signalling Pathway and its Interplay with other Parkinson's Disease Components

Development of better approaches to study PINK1 structure (joint application with University Medical Centre Goettingen and University of Frankfurt)

Novel Genome Integrity Pathways that Regulate DNA Replication Termination in Metazoa

Research output

Mechanism of human PINK1 activation at the TOM complex by reconstitution

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MRC PPU DNA Sequencing and Services

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