Abstract
Protein S-nitrosylation is known to regulate enzymatic function. Here, we report that nitric oxide (NO)-related species can contribute to Alzheimer’s disease (AD) by S-nitrosylating the lysosomal protease cathepsin B (forming SNO-CTSB), thereby inhibiting CTSB activity. This posttranslational modification inhibited autophagic flux, increased autolysosomal vesicles, and led to accumulation of protein aggregates. CA-074Me, a CTSB chemical inhibitor, also inhibited autophagic flux and resulted in accumulation of protein aggregates similar to the effect of SNO-CTSB. Inhibition of CTSB activity also induced caspase-dependent neuronal apoptosis in mouse cerebrocortical cultures. To examine which cysteine residue(s) in CTSB are S-nitrosylated, we mutated candidate cysteines and found that three cysteines were susceptible to S-nitrosylation. Finally, we observed an increase in SNO-CTSB in both 5XFAD transgenic mouse and flash-frozen postmortem human AD brains. These results suggest that S-nitrosylation of CTSB inhibits enzymatic activity, blocks autophagic flux, and thus contributes to AD pathogenesis.
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Data availability
Correspondence and requests for materials should be addressed to YHK or SAL.
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Acknowledgements
We thank Eliezer Masliah (UC San Diego/NIA) for providing human brain tissues. This work was supported in part by NIH grants R35 AG071734, R01 NS086890, R01 DA048882, DP1 DA041722, RF1 AG057409, and R01 AG056259 (to SAL), R01 AG061845, RF1 NS123298, and R61 NS122098 (to TN), and National Research Foundation of Korea (NRF) grants NRF-2017M3C7A1028945, NRF-2018R1D1A1B07049746, and NRF-2021R1A2C2008234 (to YHK).
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YHK and SAL conceived the project and designed the experiments. KRK, EJC, JWE, SSO, TN, and CKO performed experiments. KRK, TN, CKO, SAL and YHK analyzed and interpreted results. KRK, TN, CKO, YHK, and SAL wrote the manuscript.
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The authors declare that YHK is a shareholder of Zincure Corp., and that KRK is currently employed by Zincure Corp. SAL is a scientific founder of Adamas Pharmaceuticals, Inc., EuMentis Therapeutics, Inc., and InflaMED Therapeutics, LLC.
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Our studies did not include human participants or human data. Archived human brain samples were analyzed with institutional permission under the state of California and NIH guidelines. Informed consent was obtained according to procedures approved by Institutional Review Boards at the University of California, San Diego, School of Medicine, and The Scripps Research Institute. All animal experimental procedures were approved by the Animal Care and Use Committee of Sejong University and were conducted following the guidelines of the Care and Use of Laboratory Animals.
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Kim, KR., Cho, EJ., Eom, JW. et al. S-Nitrosylation of cathepsin B affects autophagic flux and accumulation of protein aggregates in neurodegenerative disorders. Cell Death Differ 29, 2137–2150 (2022). https://doi.org/10.1038/s41418-022-01004-0
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DOI: https://doi.org/10.1038/s41418-022-01004-0