Mechanism of human PINK1 activation at the TOM complex by reconstitution

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:10.1101/2023.12.23.573181

Mechanism of human PINK1 activation at the TOM complex by reconstitution

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 earlyonset Parkinson’s disease (PD). Stabilisation of PINK1 at the Translocase of Outer Membrane (TOM) complex of damaged mitochondria is a critical step for its activation. To date the mechanism of how PINK1 is activated in the TOM complex is unclear. Herein we report coexpression 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 towards 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 modelling 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 molecular findings will aid in the development of small molecule activators of PINK1 as a therapeutic strategy for PD.

Original language	English
Publisher	BioRxiv
Number of pages	35
DOIs	https://doi.org/10.1101/2023.12.23.573181
Publication status	Published - 23 Dec 2023

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Mechanism of human PINK1 activation at the TOM complex in a reconstituted system
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. (Lead / Corresponding author), Jun 2024, In: Science Advances. 10, 23, 16 p., eadn7191.
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title = "Mechanism of human PINK1 activation at the TOM complex by reconstitution",

abstract = "Loss of function mutations in PTEN-induced kinase 1 (PINK1) are a frequent cause of earlyonset Parkinson{\textquoteright}s disease (PD). Stabilisation of PINK1 at the Translocase of Outer Membrane (TOM) complex of damaged mitochondria is a critical step for its activation. To date the mechanism of how PINK1 is activated in the TOM complex is unclear. Herein we report coexpression 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 towards 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 modelling 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 molecular findings will aid in the development of small molecule activators of PINK1 as a therapeutic strategy for PD. ",

author = "Raimi, {Olawale G.} and Hina Ojha and Kenneth Ehses and Verena Dederer and Lange, {Sven M.} and {Polo Rivera}, Cristian 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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AU - Raimi, Olawale G.

AU - Ojha, Hina

AU - Ehses, Kenneth

AU - Dederer, Verena

AU - Lange, Sven M.

AU - Polo Rivera, Cristian

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.

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N2 - Loss of function mutations in PTEN-induced kinase 1 (PINK1) are a frequent cause of earlyonset Parkinson’s disease (PD). Stabilisation of PINK1 at the Translocase of Outer Membrane (TOM) complex of damaged mitochondria is a critical step for its activation. To date the mechanism of how PINK1 is activated in the TOM complex is unclear. Herein we report coexpression 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 towards 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 modelling 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 molecular 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 earlyonset Parkinson’s disease (PD). Stabilisation of PINK1 at the Translocase of Outer Membrane (TOM) complex of damaged mitochondria is a critical step for its activation. To date the mechanism of how PINK1 is activated in the TOM complex is unclear. Herein we report coexpression 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 towards 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 modelling 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 molecular findings will aid in the development of small molecule activators of PINK1 as a therapeutic strategy for PD.

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BT - Mechanism of human PINK1 activation at the TOM complex by reconstitution

PB - BioRxiv

ER -

Mechanism of human PINK1 activation at the TOM complex by reconstitution

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

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