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The N‐terminal His‐tag affects the triglyceride lipase activity of hormone‐sensitive lipase in testis
Author(s) -
Wang Feng,
Ren XiaoFang,
Chen Zheng,
Li XiaoLong,
Zhu HaiJing,
Li Sen,
Ou XiangHong,
Zhang Cheng,
Zhang FeiXiong,
Zhu BaoChang
Publication year - 2019
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.28643
Subject(s) - hormone sensitive lipase , lipase , triglyceride , cholesteryl ester , chemistry , medicine , monoacylglycerol lipase , triglyceride lipase , enzyme assay , endocrinology , lipoprotein lipase , biology , enzyme , biochemistry , cholesterol , lipoprotein , endocannabinoid system , receptor
The sterility of hormone‐sensitive lipase (HSL) knockout mice clearly shows the link between lipid metabolism and spermatogenesis. However, which substrate or product of this multifunctional lipase affects spermatogenesis is unclear. We found that an HSL protein with a His‐tag at the N‐terminus 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‐HSL) were produced on a background of HSL deficiency ( HSL −/− h ). As a result, HSL −/− h testis has an 8.65‐fold higher CE activity than wild‐type testis but a twofold higher TG activity than wild‐type testis. To compare His‐HSL and wild‐type HSL in vitro and in vivo, we confirmed that the His‐tag significantly suppressed HSL TG activity. From our results, we believe that TG activity was affected by the His‐tag insertion, but CE activity was not influenced. Furthermore, the His‐tag protected HSL from binding to the inhibitor BAY. From our study, TG activity and BAY binding sites were affected by N‐terminal His‐tag insertion.