通过抑制谷氨酰胺酶（GAC）的肿瘤代谢是打断肿瘤进展的一种有前途的策略。然而，关于GAC乙酰化的机制仍然大多数不为人知。采用线粒体蛋白质分离和谷氨酰胺酶活性检测来检查GAC活性; RT-qPCR、western blot、球形成、ALDH活性和肿瘤起始实验来评价细胞干性的变化; 构建Co-IP和挽救实验以探索潜在机制。在本研究中，我们证明GAC乙酰化是一种重要的后翻译修饰，可以抑制胶质瘤中的GAC活性。我们确定GAC被类II去乙酰化酶HDAC4去乙酰化。GAC乙酰化促进了GAC和SIRT5之间的相互作用，从而促进了GAC的泛素化并抑制了GAC的活性。此外，GAC过表达抑制了胶质瘤细胞的干性，这可以通过去乙酰化GAC来挽救。我们的研究发现了一种新的GAC乙酰化和泛素化调节机制，参与胶质瘤干性。版权所有©Bentham Science Publishers; 如有任何疑问，请发送电子邮件至epub@benthamscience.net。

Inhibiting cancer metabolism via glutaminase (GAC) is a promising strategy to disrupt tumor progression. However, the mechanism regarding GAC acetylation remains largely unknown.Mitochondrial protein isolation and glutaminase activity assay were used to examine GAC activity; RT-qPCR, western blot, sphere-formation, ALDH activity and tumor-initiating assays were performed to evaluate the alteration of cell stemness; Co-IP and rescuing experiments were constructed to explore the underlying mechanisms.In this study, we demonstrated that GAC acetylation was a vital post-translational modification that inhibits GAC activity in glioma. We identified that GAC was deacetylated by HDAC4, a class II deacetylase. GAC acetylation stimulated the interaction between GAC and SIRT5, therefore promoting GAC ubiquitination and inhibiting GAC activity. Furthermore, GAC overexpression suppressed the stemness of glioma cells, which was rescued by deacetylation of GAC.Our findings reveal a novel mechanism of GAC regulation by acetylation and ubiquitination that participates in glioma stemness.Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.