Identification of an autoinhibitory, mitophagy-inducing peptide derived from the transmembrane domain of USP30
Open accessAbstract
The mitochondrial-anchored deubiquitinating enzyme USP30 (ubiquitin specific peptidase 30) antagonizes PRKN/parkin-mediated mitophagy, making it a potential target for treating Parkinson disease. However, few inhibitors targeting USP30 have been reported. Here, we report a novel peptide (Q14) derived from the transmembrane (TM) domain of USP30 that can target mitochondrial-anchored USP30 directly and increase mitophagy through two intriguing and distinct mechanisms: a novel autoinhibition mechanism in USP30 and accelerated autophagosome formation via the LC3-interacting region (LIR) of the Q14 peptide. We identified the potential binding sites between the Q14 peptide and USP30 and postulated that an allosteric autoinhibition mechanism regulates USP30 activity. Furthermore, the LIR motif in the Q14 peptide offers additional binding with LC3 and accelerated autophagosome formation. The two mechanisms synergistically enhance mitophagy. Our work provides novel insight and direction to the design of inhibitors for USP30 or other deubiquitinating enzymes (DUBs).Abbreviations: 3-MA: 3-methyladenine; ATTEC: autophagosome-tethering compound; BafA1: bafilomycin A1; BNIP3: BCL2 interacting protein 3; BNIP3L/NIX: BCL2 interacting protein 3 like; CCCP: carbonyl cyanide m-chlorophenyl hydrazone; DMSO: dimethyl sulfoxide; FP: fluorescence polarization; FUNDC1: FUN14 domain containing 1; HCQ: hydroxychloroquine; LIR: LC3-interacting region; MST: microscale thermophoresis; mtDNA: mitochondrial DNA; mtPA-GFP: mitochondria-targeted photoactive fluorescence protein; OMM: outer mitochondrial membrane; PINK1: PTEN induced kinase 1; PRKN/parkin: parkin RBR E3 ubiquitin protein ligase; Rap: rapamycin; SA: streptavidin; TM: transmembrane; Ub: ubiquitin; Ub-AMC: Ub-7-amido-4-methylcoumarin; UPS: ubiquitin-protease system; USP: ubiquitin specific peptidase; USP30: ubiquitin specific peptidase 30.
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Q14 (231) is a novel peptide capable of traversing the cell membrane and binding to mitochondria-anchored USP30, inhibiting deubiquitination and degradation of mitochondrial proteins; it directly binds LC3 via its LIR domain, connecting USP30 and mitochondria to the autophagic membrane in an ATTEC-dependent manner, enhancing autophagic vesicle formation and promoting mitochondrial autophagy.
“In 2021, Li et al. reported a novel peptide (Q14) (231) capable of traversing the cell membrane and binding to mitochondria-anchored USP30, inhibiting deubiquitination and degradation of mitochondrial proteins (Table 4). Mechanistic investigations revealed that Q14 variably modulated the USP30 fingers domain to suppress enzymatic activity. Furthermore, Q14 directly bound LC3 via its LC3”
From Recent advances in targeting the “undruggable” proteins: from drug discovery to clinical trials (2023) · cites this paper as [540]
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- Authors
- Qin, X., Wang, R., Xu, H., Tu, L., Chen, H., Li, H., Liu, N., Wang, J., Li, S., Yin, F., Xu, N., & Li, Z.
- Affiliations
- Peking University (CN)
- Shenzhen Bay Laboratory (CN)
- Tsinghua Shenzhen International Graduate School (CN)
- Shenzhen University (CN)
- Corresponding author
- Yin, Feng
- Xu, Naihan
- Li, Zigang
- Venue
- Autophagy
- Publisher
- Informa UK Limited
- Published
- 2022-01-06
- Volume
- 18
- Issue
- 9
- Pages
- 2178-2197
- ISSN
- 1554-8627 (ISSN-L)
- 1554-8635
- PMC
- PMC9397470
- OpenAlex
- W4206675787
- Cited by
- 55
- References
- 68
- Citations by year
- 2026: 15 · 2025: 14 · 2024: 8 · 2023: 14 · 2022: 4
- FWCI
- 7.22
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Autophagy (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- China Postdoctoral Science Foundation (2021M690220)
- Natural Science Foundation of China (21778009, 21977010)
- Natural Science Foundation of Guangdong Province (2020A1515010522, 2019A1515111184, 2019A1515110487)
- National Key Research and Development Program Synthetic Biology Key Special Project of China (2018YFA0902504)
- Shenzhen Science and Technology Innovation Committee (JCYJ20170817161908969, JCYJ20180507181527112, JCYJ20180508152213145, JCYJ20170817172023838, RCJC20200714114433053)
- Beijing National Laboratory of Molecular Science open grant (BNLMS20160112)
- Shenzhen-Hong Kong Institute of Brain Science-Shenzhen Fundamental Research Institutions grant (2019SHIBS0004)
- Topics
- Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
- Health Sciences › Medicine › Neurology › Parkinson's Disease Mechanisms and Treatments
- Keywords
- USP30, mitophagy, autoinhibition, peptide inhibitors, transmembrane domain, LC3-interacting region, autophagosome formation, deubiquitinase, Parkinson's disease, allosteric regulation, mitochondrial quality control, peptide design, binding site identification
- MeSH terms
- Autophagy, Carbonyl Cyanide m-Chlorophenyl Hydrazone, DNA, Mitochondrial, Proto-Oncogene Proteins c-bcl-2, Ubiquitin, Ubiquitin-Protein Ligases, Apoptosis Regulatory Proteins, Mitophagy, Ubiquitin-Specific Proteases
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