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It also has \u03b2-lyase activity toward cystine, generating cysteine persulfide (Cys-SSH). The chemical reactivity of Cys-SSH is thought to be involved in the catalytic activity of particular proteins via protein polysulfidation, the formation of -S-(S)n-H on their reactive cysteine residues. The Cys136\/171 residues of CSE have been proposed to be redox-sensitive residues. Herein, we investigated whether CSE polysulfidation occurs at Cys136\/171 during cystine metabolism. Transfection of wild-type CSE into COS-7 cells resulted in increased intracellular Cys-SSH production, which was significantly increased when Cys136Val or Cys136\/171Val CSE mutants were transfected, instead of the wild-type enzyme. A biotin-polyethylene glycol-conjugated maleimide capture assay revealed that CSE polysulfidation occurs at Cys136 during cystine metabolism. In vitro incubation of CSE with CSE-enzymatically synthesized Cys-SSH resulted in the inhibition of Cys-SSH production. In contrast, the mutant CSEs (Cys136Val and Cys136\/171Val) proved resistant to inhibition. The Cys-SSH-producing CSE activity of Cys136\/171Val CSE was higher than that of the wild-type enzyme. Meanwhile, the cysteine-producing CSE activity of this mutant was equivalent to that of the wild-type enzyme. It is assumed that Cys-SSH-producing CSE activity could be auto-inactivated via the polysulfidation of the enzyme during cystine metabolism. Thus, the polysulfidation of CSE at the Cys136 residue may be an integral feature of cystine metabolism, which functions to down-regulate Cys-SSH synthesis by the enzyme.<\/jats:p>","DOI":"10.3390\/ijms24129982","type":"journal-article","created":{"date-parts":[[2023,6,12]],"date-time":"2023-06-12T03:02:27Z","timestamp":1686538947000},"page":"9982","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Cystathionine \u03b3-Lyase Self-Inactivates by Polysulfidation during Cystine Metabolism"],"prefix":"10.3390","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2034-2799","authenticated-orcid":false,"given":"Shoma","family":"Araki","sequence":"first","affiliation":[{"name":"Department of Pharmacology, Showa Pharmaceutical University, Machida 194-8543, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6691-2569","authenticated-orcid":false,"given":"Tsuyoshi","family":"Takata","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Showa Pharmaceutical University, Machida 194-8543, Japan"},{"name":"South Bend Campus, Indiana University School of Medicine, South Bend, IN 46617, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katsuhiko","family":"Ono","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Graduate School of Medical Sciences, Kumamoto University, Kumamoto 860-8556, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7949-7309","authenticated-orcid":false,"given":"Tomohiro","family":"Sawa","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Graduate School of Medical Sciences, Kumamoto University, Kumamoto 860-8556, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3094-8543","authenticated-orcid":false,"given":"Shingo","family":"Kasamatsu","sequence":"additional","affiliation":[{"name":"Department of Biological Chemistry, Graduate School of Science, Osaka Metropolitan University, Sakai 599-8531, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hideshi","family":"Ihara","sequence":"additional","affiliation":[{"name":"Department of Biological Chemistry, Graduate School of Science, Osaka Metropolitan University, Sakai 599-8531, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoshito","family":"Kumagai","sequence":"additional","affiliation":[{"name":"Graduate School of Pharmaceutical Sciences, Kyusyu University, Fukuoka 812-8582, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0623-1710","authenticated-orcid":false,"given":"Takaaki","family":"Akaike","sequence":"additional","affiliation":[{"name":"Department of Environmental Medicine and Molecular Toxicology, Graduate School of Medicine, Tohoku University, Sendai 980-8575, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2232-0091","authenticated-orcid":false,"given":"Yasuo","family":"Watanabe","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Showa Pharmaceutical University, Machida 194-8543, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1526-3351","authenticated-orcid":false,"given":"Yukihiro","family":"Tsuchiya","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Showa Pharmaceutical University, Machida 194-8543, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,6,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1111\/bph.14446","article-title":"Regulators of the transsulfuration pathway","volume":"176","author":"Sbodio","year":"2019","journal-title":"Br. 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