Site-specific cleavage of acetoacetyl-CoA synthetase by legumain
Site-specific cleavage of acetoacetyl-CoA synthetase by legumain
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DOI：
10.1002/1873-3468.12197
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发表时间：
2016-06-01
期刊：
影响因子：
3.5
通讯作者：
Fukui, Tetsuya
中科院分区：
文献类型：
--
作者：
Hasegawa, Shinya;Inoue, Daiki;Fukui, Tetsuya
乙酰乙酰-辅酶A合成酶(AACS)是一种酮体利用酶，负责胆固醇和脂肪酸的合成。我们先前已经证明，AACS被一种溶酶体天冬酰胺内肽酶--豆蛋白切割。在本研究中，我们试图确定AAC的切割位点。AACs的诱变分析表明，Asn547是AACs在小鼠肝脏中的特异性切割位点。裂解形式的AACS(1-547)失去了将乙酰乙酸酯转化为乙酰乙酰-辅酶A的能力。此外，基于流体力学的基因转导表明，AAC(1-547)的过表达增加了小窝的主要成分小凹-1的蛋白表达。这些结果表明，豆类蛋白对AAC的切割对于酶活性的调节和功能获得的改变是至关重要的。
Acetoacetyl-CoA synthetase (AACS) is a ketone body-utilizing enzyme and is responsible for the synthesis of cholesterol and fatty acids. We have previously shown that AACS is cleaved by legumain, a lysosomal asparaginyl endopeptidase. In this study, we attempted to determine the cleavage site of AACS. Mutagenesis analysis of AACS revealed that Asn547 is the specific cleavage site of AACS in mouse livers. The cleaved form of AACS (1-547) lost the ability to convert acetoacetate to acetoacetyl-CoA. Moreover, hydrodynamics-based gene transduction showed that overexpression of AACS (1-547) increases the protein expression of caveolin-1, the principal component of the caveolae. These results suggest that cleavage of AACS by legumain is critical for the regulation of enzymatic activity and results in gain-of-function changes.