CoChat
Sign Up
Cited by 940Parkinson's Disease Mechanisms and TreatmentsNuclear Receptors and SignalingAutophagy in Disease and Therapy

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

Open access
Kondapalli, Chandana, Kazlauskaite, Agne, Zhang, Ning, Woodroof, Helen I., Campbell, David G., Gourlay, Robert, Burchell, Lynn, Walden, Helen +5 more·2012·Open Biology
DOI 10.1098/rsob.120080PMC PMC3376738
Open this paper in CoChat — free account

Abstract

Summary: Missense mutations in PTEN-induced kinase 1 (PINK1) cause autosomal-recessive 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 Ser65. We show that human PINK1 is specifically activated by mitochondrial membrane potential (Δψm) depolarization, enabling it to phosphorylate Parkin at Ser65. We further show that phosphorylation of Parkin at Ser65 leads to marked activation of its E3 ligase activity that is prevented by mutation of Ser65 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 Δψm depolarization and suggest that PINK1 directly phosphorylates and activates Parkin. Our findings indicate that monitoring phosphorylation of Parkin at Ser65 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.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1098/rsob.120080

Cites

6 of 6 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Phosphorylation of ubiquitin and PRKN at Ser65 by PINK1 is required for activation of the PINK1-PRKN pathway.

    “Both events are required for the activation of the pathway.”

    From miRNA family miR-29 inhibits PINK1-PRKN signaling via ATG9A (2026) · cites this paper as [8]

    The Results section reports that PINK1 phosphorylates Parkin (PRKN) at Ser65 and that a non-phosphorylatable Parkin S65A mutant does not show the observed E3 ligase activation. It does not establish that PINK1 phosphorylates ubiquitin at Ser65 or that ubiquitin phosphorylation is also required for pathway activation; ubiquitin is described as a component and product of the ubiquitylation assay. (LLM, conf=0.89)

    Experimental — accuracy not yet measured

  • Upon mitochondrial depolarization, PINK1 stabilizes on the outer mitochondrial membrane and phosphorylates ubiquitin and PRKN at Ser65.

    “Upon mitochondrial depolarization, PINK1 stabilizes on the outer mitochondrial membrane, where it phosphorylates ubiquitin (Ub) and PRKN at Ser65 [5–10].”

    From miRNA family miR-29 inhibits PINK1-PRKN signaling via ATG9A (2026) · cites this paper as [8]

    The Introduction and Discussion report that depolarization stabilizes PINK1 at mitochondria and that PINK1 phosphorylates Parkin (PRKN) at Ser65. They describe relocation to the outer mitochondrial membrane as a proposal, not an established result. The Results discuss ubiquitin in ubiquitylation assays but do not show that PINK1 phosphorylates ubiquitin. (LLM, conf=0.89)

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 37

  1. [1]Muqit MM 2006 Altered cleavage and localization of PINK1 to aggresomes in the presence of proteasomal stress. J. Neurochem. 98, 156– 169 10.1111/j.1471-4159.2006.03845.x (doi:10.1111/j.1471-4159.2006.03845.x) 16805805 · DOI
  2. [3]Woodroof HI 2011 Discovery of catalytically active orthologues of the Parkinson's disease kinase PINK1: analysis of substrate specificity and impact of mutations. Open Biol. 1, 110012 10.1098/rsob.110012 (doi:10.1098/rsob.110012) 22645651 PMC3352081 · DOI
  3. [4]Clark IE Dodson MW Jiang C Cao JH Huh JR Seol JH Yoo SJ Hay BA Guo M 2006 Drosophila PINK1 is required for mitochondrial function and interacts genetically with Parkin. Nature 441, 1162– 1166 10.1038/nature04779 (doi:10.1038/nature04779) 16672981 · DOI · Paper page
  4. [5]Park J 2006 Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by Parkin. Nature 441, 1157– 1161 10.1038/nature04788 (doi:10.1038/nature04788) 16672980 · DOI · Paper page
  5. [6]Yang Y Ouyang Y Yang L Beal MF McQuibban A Vogel H Lu B 2008 PINK1 regulates mitochondrial dynamics through interaction with the fission/fusion machinery. Proc. Natl Acad. Sci. USA 105, 7070– 7075 10.1073/pnas.0711845105 (doi:10.1073/pnas.0711845105) 18443288 PMC2383971 · DOI
  6. [7]Deas E 2011 PINK1 cleavage at position A103 by the mitochondrial protease PARL. Hum. Mol. Genet. 20, 867– 879 10.1093/hmg/ddq526 (doi:10.1093/hmg/ddq526) 21138942 PMC3033179 · DOI
  7. [8]Jin SM Lazarou M Wang C Kane LA Narendra DP Youle RJ 2010 Mitochondrial membrane potential regulates PINK1 import and proteolytic destabilization by PARL. J. Cell Biol. 191, 933– 942 10.1083/jcb.201008084 (doi:10.1083/jcb.201008084) 21115803 PMC2995166 · DOI · Paper page
  8. [9]Meissner C Lorenz H Weihofen A Selkoe DJ Lemberg MK 2011 The mitochondrial intramembrane protease PARL cleaves human PINK1 to regulate PINK1 trafficking. J. Neurochem. 117, 856– 867 10.1111/j.1471-4159.2011.07253.x (doi:10.1111/j.1471-4159.2011.07253.x) 21426348 · DOI
  9. [10]Whitworth AJ Lee JR Ho VM Flick R Chowdhury R McQuibban GA 2008 Rhomboid-7 and HtrA2/Omi act in a common pathway with the Parkinson's disease factors PINK1 and Parkin. Dis. Model. Mech. 1, 168– 174 discussion 173 10.1242/dmm.000109 (doi:10.1242/dmm.000109) 19048081 PMC2562193 · DOI
  10. [11]Takatori S Ito G Iwatsubo T 2008 Cytoplasmic localization and proteasomal degradation of N-terminally cleaved form of PINK1. Neurosci. Lett. 430, 13– 17 10.1016/j.neulet.2007.10.019 (doi:10.1016/j.neulet.2007.10.019) 18031932 · DOI
  11. [15]Vives-Bauza C 2010 PINK1-dependent recruitment of Parkin to mitochondria in mitophagy. Proc. Natl Acad. Sci. USA 107, 378– 383 10.1073/pnas.0911187107 (doi:10.1073/pnas.0911187107) 19966284 PMC2806779 · DOI · Paper page
  12. [16]Kitada T Asakawa S Hattori N Matsumine H Yamamura Y Minoshima S Yokochi M Mizuno Y Shimizu N 1998 Mutations in the Parkin gene cause autosomal recessive juvenile Parkinsonism. Nature 392, 605– 608 10.1038/33416 (doi:10.1038/33416) 9560156 · DOI · Paper page
  13. [17]Exner N 2007 Loss-of-function of human PINK1 results in mitochondrial pathology and can be rescued by Parkin. J. Neurosci. 27, 12 413– 12 418 10.1523/JNEUROSCI.0719-07.2007 (doi:10.1523/JNEUROSCI.0719-07.2007) PMC6673250 17989306 · DOI
  14. [18]Abeliovich A Flint Beal M 2006 Parkinsonism genes: culprits and clues. J. Neurochem. 99, 1062– 1072 10.1111/j.1471-4159.2006.04102.x (doi:10.1111/j.1471-4159.2006.04102.x) 16836655 · DOI
  15. [19]Muqit MM Alessi DR 2009 A meeting of minds: overcoming roadblocks in the development of therapies for neurodegenerative disorders. EMBO Mol. Med. 1, 139– 141 10.1002/emmm.200900021 (doi:10.1002/emmm.200900021) 20049713 PMC3378128 · DOI
  16. [20]Plun-Favreau H 2007 The mitochondrial protease HtrA2 is regulated by Parkinson's disease-associated kinase PINK1. Nat. Cell Biol. 9, 1243– 1252 10.1038/ncb1644 (doi:10.1038/ncb1644) 17906618 · DOI
  17. [22]Wang X 2011 PINK1 and Parkin target Miro for phosphorylation and degradation to arrest mitochondrial motility. Cell. 147, 893– 906 10.1016/j.cell.2011.10.018 (doi:10.1016/j.cell.2011.10.018) 22078885 PMC3261796 · DOI
  18. [23]Weihofen A Thomas KJ Ostaszewski BL Cookson MR Selkoe DJ 2009 PINK1 forms a multiprotein complex with Miro and Milton, linking PINK1 function to mitochondrial trafficking. Biochemistry 48, 2045– 2052 10.1021/bi8019178 (doi:10.1021/bi8019178) 19152501 PMC2693257 · DOI
  19. [24]Chaugule VK Burchell L Barber KR Sidhu A Leslie SJ Shaw GS Walden H 2011 Autoregulation of Parkin activity through its ubiquitin-like domain. EMBO J. 30, 2853– 2867 10.1038/emboj.2011.204 (doi:10.1038/emboj.2011.204) 21694720 PMC3160258 · DOI · Paper page
  20. [25]Kim Y Park J Kim S Song S Kwon SK Lee SH Kitada T Kim JM Chung J 2008 PINK1 controls mitochondrial localization of Parkin through direct phosphorylation. Biochem. Biophys. Res. Commun. 377, 975– 980 10.1016/j.bbrc.2008.10.104 (doi:10.1016/j.bbrc.2008.10.104) 18957282 · DOI
  21. [26]Abou-Sleiman PM Muqit MM Wood NW 2006 Expanding insights of mitochondrial dysfunction in Parkinson's disease. Nat. Rev. Neurosci. 7, 207– 219 10.1038/nrn1868 (doi:10.1038/nrn1868) 16495942 · DOI · Paper page
  22. [27]Cui M Tang X Christian WV Yoon Y Tieu K 2011 Perturbations in mitochondrial dynamics induced by human mutant PINK1 can be rescued by the mitochondrial division inhibitor mdivi-1. J. Biol. Chem. 285, 11 740– 11 752 PMC2857048 20164189
  23. [28]Whitworth AJ Pallanck LJ 2009 The PINK1/Parkin pathway: a mitochondrial quality control system? J. Bioenerg. Biomembr. 41, 499– 503 10.1007/s10863-009-9253-3 (doi:10.1007/s10863-009-9253-3) 19967438 · DOI
  24. [29]Yu W Sun Y Guo S Lu B 2011 The PINK1/Parkin pathway regulates mitochondrial dynamics and function in mammalian hippocampal and dopaminergic neurons. Hum. Mol. Genet. 20, 3227– 3240 10.1093/hmg/ddr235 (doi:10.1093/hmg/ddr235) 21613270 PMC3140825 · DOI
  25. [30]Wenzel DM Lissounov A Brzovic PS Klevit RE 2011 UBCH7 reactivity profile reveals Parkin and HHARI to be RING/HECT hybrids. Nature 474, 105– 108 10.1038/nature09966 (doi:10.1038/nature09966) 21532592 PMC3444301 · DOI
  26. [31]Trempe JF Chen CX Grenier K Camacho EM Kozlov G McPherson PS Gehring K Fon EA 2009 SH3 domains from a subset of BAR proteins define a Ubl-binding domain and implicate Parkin in synaptic ubiquitination. Molecular Cell 36, 1034– 1047 10.1016/j.molcel.2009.11.021 (doi:10.1016/j.molcel.2009.11.021) 20064468 · DOI
  27. [32]Sakata E 2003 Parkin binds the Rpn10 subunit of 26S proteasomes through its ubiquitin-like domain. EMBO Rep. 4, 301– 306 10.1038/sj.embor.embor764 (doi:10.1038/sj.embor.embor764) 12634850 PMC1315892 · DOI
  28. [33]Tashiro M Okubo S Shimotakahara S Hatanaka H Yasuda H Kainosho M Yokoyama S Shindo H 2003 NMR structure of ubiquitin-like domain in PARKIN: gene product of familial Parkinson's disease. J. Biomol. NMR 25, 153– 156 10.1023/A:1022254432039 (doi:10.1023/A:1022254432039) 12652124 · DOI
  29. [34]Tomoo K Mukai Y In Y Miyagawa H Kitamura K Yamano A Shindo H Ishida T 2008 Crystal structure and molecular dynamics simulation of ubiquitin-like domain of murine Parkin. Biochim. Biophys. Acta 1784, 1059– 1067 18485927
  30. [35]Ziviani E Tao RN Whitworth AJ 2010 Drosophila Parkin requires PINK1 for mitochondrial translocation and ubiquitinates mitofusin. Proc. Natl Acad. Sci. USA 107, 5018– 5023 10.1073/pnas.0913485107 (doi:10.1073/pnas.0913485107) 20194754 PMC2841909 · DOI · Paper page
  31. [36]Shin JH Ko HS Kang H Lee Y Lee YI Pletinkova O Troconso JC Dawson VL Dawson TM 2011 PARIS (ZNF746) repression of PGC-1alpha contributes to neurodegeneration in Parkinson's disease. Cell 144, 689– 702 10.1016/j.cell.2011.02.010 (doi:10.1016/j.cell.2011.02.010) 21376232 PMC3063894 · DOI · Paper page
  32. [38]Xiong H 2009 Parkin, PINK1, and DJ-1 form a ubiquitin E3 ligase complex promoting unfolded protein degradation. J. Clin. Invest. 119, 650– 660 10.1172/JCI37617 (doi:10.1172/JCI37617) 19229105 PMC2648688 · DOI
  33. [39]Sha D Chin LS Li L 2010 Phosphorylation of Parkin by Parkinson disease-linked kinase PINK1 activates Parkin E3 ligase function and NF-kappaB signaling. Hum. Mol. Genet. 19, 352– 363 10.1093/hmg/ddp501 (doi:10.1093/hmg/ddp501) 19880420 PMC2796895 · DOI
  34. [40]Avraham E Rott R Liani E Szargel R Engelender S 2007 Phosphorylation of Parkin by the cyclin-dependent kinase 5 at the linker region modulates its ubiquitin-ligase activity and aggregation. J. Biol. Chem. 282, 12 842– 12 850 10.1074/jbc.M608243200 (doi:10.1074/jbc.M608243200) 17327227 · DOI
  35. [41]Reed SE Staley EM Mayginnes JP Pintel DJ Tullis GE 2006 Transfection of mammalian cells using linear polyethylenimine is a simple and effective means of producing recombinant adeno-associated virus vectors. J. Virol. Meth. 138, 85– 98 10.1016/j.jviromet.2006.07.024 (doi:10.1016/j.jviromet.2006.07.024) 16950522 · DOI
  36. [42]Campbell DG Morrice NA 2002 Identification of protein phosphorylation sites by a combination of mass spectrometry and solid phase Edman sequencing. J. Biomol. Techn. 13, 119– 130 PMC2279855 19498976
  37. [43]Williamson BL Marchese J Morrice NA 2006 Automated identification and quantification of protein phosphorylation sites by LC/MS on a hybrid triple quadrupole linear ion trap mass spectrometer. Mol. Cell. Proteom. 5, 337– 346 10.1074/mcp.M500210-MCP200 (doi:10.1074/mcp.M500210-MCP200) 16260767 · DOI

Cited by

Sorted by citations

Authors
Kondapalli, C., Kazlauskaite, A., Zhang, N., Woodroof, H. I., Campbell, D. G., Gourlay, R., Burchell, L., Walden, H., Macartney, T. J., Deak, M., Knebel, A., Alessi, D. R., & Muqit, M. M. K.
Affiliations
  • University of Dundee (GB)
  • The Honourable Society of Lincoln's Inn (GB)
  • University of Dundee 1 , ,
Corresponding author
Muqit, Miratul M. K.
ORCID
Venue
Open Biology
Publisher
The Royal Society
Published
2012-01-01
Volume
2
Issue
5
Pages
120080-120080
ISSN
2046-2441 (ISSN-L)
DOI
10.1098/rsob.120080
PMC
PMC3376738
OpenAlex
W2140654974
Cited by
940
References
6
Citations by year
2026: 48 · 2025: 58 · 2024: 90 · 2023: 57 · 2022: 85 · 2021: 96 · 2020: 90 · 2019: 57 · 2018: 70 · 2017: 55 · 2016: 60 · 2015: 72 · 2014: 58 · 2013: 37 · 2012: 6
FWCI
36.08
Type
journal article
Language
en
Open access
Licence
Locations
  • Open Biology (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • ENLIGHTEN (Jurnal Bimbingan dan Konseling Islam) (acceptedVersion)
  • Discovery Research Portal (University of Dundee) (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Wellcome Trust (089698)
  • Parkinson's UK (H-0901, H-1101)
  • Medical Research Council (MC_G1000735)
Topics
  • Health Sciences › Medicine › Neurology › Parkinson's Disease Mechanisms and Treatments
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Nuclear Receptors and Signaling
  • Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
Keywords
PINK1, Parkin, Parkinson's disease, mitochondrial depolarization, biomarkers
MeSH terms
PTEN-Induced Putative Kinase, Animals, Carbonyl Cyanide m-Chlorophenyl Hydrazone, Enzyme Activation, Humans, Parkinson Disease, Phosphorylation, Phosphoserine, Phosphothreonine, Protein Kinases, Protein Processing, Post-Translational, Recombinant Fusion Proteins, Tribolium, Protein Structure, Tertiary, Insect Proteins, RNA Interference, RNA, Small Interfering, Ubiquitin-Protein Ligases, Membrane Potential, Mitochondrial, Receptor-Like Protein Tyrosine Phosphatases, Class 2, Protein Stability, HEK293 Cells

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review