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However, which substrate or product of this multifunctional lipase affects spermatogenesis is unclear. We found that an HSL protein with a His\u2010tag at the N\u2010terminus preserved the normal hydrolase activity of cholesteryl ester (CE) but the triglyceride lipase (TG) activity significantly decreased in vitro. Therefore, mice with this functionally incomplete HSL (His\u2010HSL) were produced on a background of HSL deficiency (<jats:italic>HSL<jats:sup>\u2212\/\u2212<\/jats:sup>h<\/jats:italic>). As a result, <jats:italic>HSL<jats:sup>\u2212\/\u2212<\/jats:sup>h<\/jats:italic> testis has an 8.65\u2010fold higher CE activity than wild\u2010type testis but a twofold higher TG activity than wild\u2010type testis. To compare His\u2010HSL and wild\u2010type HSL in vitro and in vivo, we confirmed that the His\u2010tag significantly suppressed HSL TG activity. From our results, we believe that TG activity was affected by the His\u2010tag insertion, but CE activity was not influenced. Furthermore, the His\u2010tag protected HSL from binding to the inhibitor BAY. From our study, TG activity and BAY binding sites were affected by N\u2010terminal His\u2010tag insertion.<\/jats:p>","DOI":"10.1002\/jcb.28643","type":"journal-article","created":{"date-parts":[[2019,4,2]],"date-time":"2019-04-02T06:24:55Z","timestamp":1554186295000},"page":"13706-13716","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["The N\u2010terminal His\u2010tag affects the triglyceride lipase activity of hormone\u2010sensitive lipase in testis"],"prefix":"10.1002","volume":"120","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4591-3888","authenticated-orcid":false,"given":"Feng","family":"Wang","sequence":"first","affiliation":[{"name":"College of Life Sciences Capital Normal University Beijing China"},{"name":"Fertility Preservation Lab, Reproductive Medicine Center Guangdong Second Provincial General Hospital Guangzhou 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