Ubiquitome analysis reveals the involvement of lysine ubiquitination in the spermatogenesis process of adult buffalo (Bubalus bubalis) testis
Ubiquitome analysis reveals the involvement of lysine ubiquitination in the spermatogenesis process of adult buffalo (Bubalus bubalis) testis
复制标题
泛素组分析揭示赖氨酸泛素化参与成年水牛（Bubalus bubalis）睾丸的精子发生过程
DOI：
10.1042/bsr20193537
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发表时间：
2020-06-16
影响因子：
4
通讯作者：
Zhang, Ming
中科院分区：
文献类型：
--
作者：
Huang, Yu-li;Zhang, Peng-fei;Zhang, Ming
蛋白质泛素化是一种重要的、保守的翻译后修饰，在真核生物的许多生物学过程中起着重要的调控作用。虽然在我们以前的研究中已经在布法罗睾丸中发现了几种泛素化蛋白，但迄今为止还没有大规模的布法罗睾丸泛素化组的研究报道。在本研究中，我们首先确定了一个全球性的成人布法罗睾丸赖氨酸泛素化的分析，使用高灵敏度的LC-MS/MS结合免疫亲和富集的泛素化肽。在成年布法罗睾丸组织的262个蛋白质中共鉴定出422个赖氨酸泛素化位点。生物信息学分析表明，泛素化蛋白参与了多种生物学过程和不同的亚细胞定位。京都基因和基因组百科全书（KEGG）途径和蛋白质相互作用网络分析表明，睾丸中蛋白质泛素化对蛋白酶体、糖酵解/代谢产物生成和缝隙连接途径有调节作用。此外，SpermatogenesisOnline数据库中还发现了44个可能参与精子发生的泛素化蛋白，其中HSPA 2和UCHL 1的泛素化已被免疫沉淀/Western blot证实。综上所述，这些数据首次提供了布法罗睾丸泛素化的整体视图，并为探索哺乳动物睾丸赖氨酸泛素化的生理作用，特别是精子发生的重要资源。
Protein ubiquitination, a major and conserved post-translational modification, is known to play a critical regulatory role in many biological processes in eukaryotes. Although several ubiquitinated proteins have been found in buffalo (Bubalus bubalis) testis in our previous studies, large-scale profiling of buffalo testis ubiquitome has not been reported to date. In the present study, we first identified a global profiling of lysine ubiquitination of adult buffalo testis using a highly sensitive LC-MS/MS coupled with immune-affinity enrichment of ubiquitinated peptides. In total, 422 lysine ubiquitination sites were identified in 262 proteins in adult buffalo testis tissue. Bioinformatics analysis showed that the ubiquitinated proteins are involved in a variety of biological processes and diverse subcellular localizations. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and protein interaction network analysis indicated that proteasome, glycolysis/gluconeogenesis and gap junction pathways are modulated by protein ubiquitination in testis. Besides, 44 ubiquitinated proteins may involve in spermatogenesis according to the SpermatogenesisOnline database, of which, the ubiquitination of HSPA2 and UCHL1 were confirmed by Immunoprecipitation (IP)/Western blot analysis. Taken together, these data provide a global view of ubiquitome in buffalo testis for the first time, and serve as an important resource for exploring the physiological role especially spermatogenesis of lysine ubiquitination in testis in mammals.