PINK1 is activated by mitochondrial membrane potential depolarization and stimulates Parkin E3 ligase activity by phosphorylating Serine 65

Chandana Kondapalli, Agne Kazlauskaite, Ning Zhang, Helen I. Woodroof, David G. Campbell, Robert Gourlay, Lynn Burchell, Helen Walden, Thomas J. MacArtney, Maria Deak, Axel Knebel, Dario R. Alessi, Miratul M. K. Muqit (Lead / Corresponding author)

    Research output: Contribution to journalArticle

    336 Citations (Scopus)

    Abstract

    Missense mutations in PTEN-induced kinase 1 (PINK1) cause autosomalrecessive inherited Parkinson's disease (PD). We have exploited our recent discovery that recombinant insect PINK1 is catalytically active to test whether PINK1 directly phosphorylates 15 proteins encoded by PD-associated genes as well as proteins reported to bind PINK1. We have discovered that insect PINK1 efficiently phosphorylates only one of these proteins, namely the E3 ligase Parkin. We have mapped the phosphorylation site to a highly conserved residue within the Ubl domain of Parkin at Ser . We show that human PINK1 is specifically activated by mitochondrial membrane potential (? ?m) depolarization, enabling it to phosphorylate Parkin at Ser . We further show that phosphorylation of Parkin at Ser leads to marked activation of its E3 ligase activity that is prevented by mutation of Ser or inactivation of PINK1. We provide evidence that once activated, PINK1 autophosphorylates at several residues, including Thr257, which is accompanied by an electrophoretic mobility band-shift. These results provide the first evidence that PINK1 is activated following Dcm depolarization and suggest that PINK1 directly phosphorylates and activates Parkin. Our findings indicate that monitoring phosphorylation of Parkin at Ser and/or PINK1 at Thr257 represent the first biomarkers for examining activity of the PINK1-Parkin signalling pathway in vivo. Our findings also suggest that small molecule activators of Parkin that mimic the effect of PINK1 phosphorylation may confer therapeutic benefit for PD.
    Original languageEnglish
    Article number120080
    JournalOpen Biology
    Volume2
    DOIs
    Publication statusPublished - 2012

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    Ubiquitin-Protein Ligases
    Mitochondrial Membrane Potential
    Depolarization
    Serine
    Phosphotransferases
    Membranes
    Phosphorylation
    Parkinson Disease
    Insects
    Electrophoretic mobility
    Proteins
    Biomarkers
    Missense Mutation

    Cite this

    Kondapalli, Chandana ; Kazlauskaite, Agne ; Zhang, Ning ; Woodroof, Helen I. ; Campbell, David G. ; Gourlay, Robert ; Burchell, Lynn ; Walden, Helen ; MacArtney, Thomas J. ; Deak, Maria ; Knebel, Axel ; Alessi, Dario R. ; Muqit, Miratul M. K. / PINK1 is activated by mitochondrial membrane potential depolarization and stimulates Parkin E3 ligase activity by phosphorylating Serine 65. In: Open Biology. 2012 ; Vol. 2.
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    title = "PINK1 is activated by mitochondrial membrane potential depolarization and stimulates Parkin E3 ligase activity by phosphorylating Serine 65",
    abstract = "Missense mutations in PTEN-induced kinase 1 (PINK1) cause autosomalrecessive inherited Parkinson's disease (PD). We have exploited our recent discovery that recombinant insect PINK1 is catalytically active to test whether PINK1 directly phosphorylates 15 proteins encoded by PD-associated genes as well as proteins reported to bind PINK1. We have discovered that insect PINK1 efficiently phosphorylates only one of these proteins, namely the E3 ligase Parkin. We have mapped the phosphorylation site to a highly conserved residue within the Ubl domain of Parkin at Ser . We show that human PINK1 is specifically activated by mitochondrial membrane potential (? ?m) depolarization, enabling it to phosphorylate Parkin at Ser . We further show that phosphorylation of Parkin at Ser leads to marked activation of its E3 ligase activity that is prevented by mutation of Ser or inactivation of PINK1. We provide evidence that once activated, PINK1 autophosphorylates at several residues, including Thr257, which is accompanied by an electrophoretic mobility band-shift. These results provide the first evidence that PINK1 is activated following Dcm depolarization and suggest that PINK1 directly phosphorylates and activates Parkin. Our findings indicate that monitoring phosphorylation of Parkin at Ser and/or PINK1 at Thr257 represent the first biomarkers for examining activity of the PINK1-Parkin signalling pathway in vivo. Our findings also suggest that small molecule activators of Parkin that mimic the effect of PINK1 phosphorylation may confer therapeutic benefit for PD.",
    author = "Chandana Kondapalli and Agne Kazlauskaite and Ning Zhang and Woodroof, {Helen I.} and Campbell, {David G.} and Robert Gourlay and Lynn Burchell and Helen Walden and MacArtney, {Thomas J.} and Maria Deak and Axel Knebel and Alessi, {Dario R.} and Muqit, {Miratul M. K.}",
    note = "Copyright 2012 Elsevier B.V., All rights reserved.",
    year = "2012",
    doi = "10.1098/rsob.120080",
    language = "English",
    volume = "2",
    journal = "Open Biology",
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    PINK1 is activated by mitochondrial membrane potential depolarization and stimulates Parkin E3 ligase activity by phosphorylating Serine 65. / Kondapalli, Chandana; Kazlauskaite, Agne; Zhang, Ning; Woodroof, Helen I.; Campbell, David G.; Gourlay, Robert; Burchell, Lynn; Walden, Helen; MacArtney, Thomas J.; Deak, Maria; Knebel, Axel; Alessi, Dario R.; Muqit, Miratul M. K. (Lead / Corresponding author).

    In: Open Biology, Vol. 2, 120080, 2012.

    Research output: Contribution to journalArticle

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    AU - Kondapalli, Chandana

    AU - Kazlauskaite, Agne

    AU - Zhang, Ning

    AU - Woodroof, Helen I.

    AU - Campbell, David G.

    AU - Gourlay, Robert

    AU - Burchell, Lynn

    AU - Walden, Helen

    AU - MacArtney, Thomas J.

    AU - Deak, Maria

    AU - Knebel, Axel

    AU - Alessi, Dario R.

    AU - Muqit, Miratul M. K.

    N1 - Copyright 2012 Elsevier B.V., All rights reserved.

    PY - 2012

    Y1 - 2012

    N2 - Missense mutations in PTEN-induced kinase 1 (PINK1) cause autosomalrecessive inherited Parkinson's disease (PD). We have exploited our recent discovery that recombinant insect PINK1 is catalytically active to test whether PINK1 directly phosphorylates 15 proteins encoded by PD-associated genes as well as proteins reported to bind PINK1. We have discovered that insect PINK1 efficiently phosphorylates only one of these proteins, namely the E3 ligase Parkin. We have mapped the phosphorylation site to a highly conserved residue within the Ubl domain of Parkin at Ser . We show that human PINK1 is specifically activated by mitochondrial membrane potential (? ?m) depolarization, enabling it to phosphorylate Parkin at Ser . We further show that phosphorylation of Parkin at Ser leads to marked activation of its E3 ligase activity that is prevented by mutation of Ser or inactivation of PINK1. We provide evidence that once activated, PINK1 autophosphorylates at several residues, including Thr257, which is accompanied by an electrophoretic mobility band-shift. These results provide the first evidence that PINK1 is activated following Dcm depolarization and suggest that PINK1 directly phosphorylates and activates Parkin. Our findings indicate that monitoring phosphorylation of Parkin at Ser and/or PINK1 at Thr257 represent the first biomarkers for examining activity of the PINK1-Parkin signalling pathway in vivo. Our findings also suggest that small molecule activators of Parkin that mimic the effect of PINK1 phosphorylation may confer therapeutic benefit for PD.

    AB - Missense mutations in PTEN-induced kinase 1 (PINK1) cause autosomalrecessive inherited Parkinson's disease (PD). We have exploited our recent discovery that recombinant insect PINK1 is catalytically active to test whether PINK1 directly phosphorylates 15 proteins encoded by PD-associated genes as well as proteins reported to bind PINK1. We have discovered that insect PINK1 efficiently phosphorylates only one of these proteins, namely the E3 ligase Parkin. We have mapped the phosphorylation site to a highly conserved residue within the Ubl domain of Parkin at Ser . We show that human PINK1 is specifically activated by mitochondrial membrane potential (? ?m) depolarization, enabling it to phosphorylate Parkin at Ser . We further show that phosphorylation of Parkin at Ser leads to marked activation of its E3 ligase activity that is prevented by mutation of Ser or inactivation of PINK1. We provide evidence that once activated, PINK1 autophosphorylates at several residues, including Thr257, which is accompanied by an electrophoretic mobility band-shift. These results provide the first evidence that PINK1 is activated following Dcm depolarization and suggest that PINK1 directly phosphorylates and activates Parkin. Our findings indicate that monitoring phosphorylation of Parkin at Ser and/or PINK1 at Thr257 represent the first biomarkers for examining activity of the PINK1-Parkin signalling pathway in vivo. Our findings also suggest that small molecule activators of Parkin that mimic the effect of PINK1 phosphorylation may confer therapeutic benefit for PD.

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    U2 - 10.1098/rsob.120080

    DO - 10.1098/rsob.120080

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