School of Marine Science, Ningbo University, Ningbo 315020, China.
Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China.
J Proteomics. 2020 Aug 30;226:103891. doi: 10.1016/j.jprot.2020.103891. Epub 2020 Jul 4.
Lysine crotonylation (Kcr) is a recently-discovered type of post-translational modification. Although Kcr has been reported in many species, little is known about this process in crustaceans. In this study, pan anti-lysine crotonylation antibody enrichment and high-resolution liquid chromatogram-mass spectrometry analysis were employed to characterize Kcr in testis of the Chinese mitten crab Eriocheir sinensis testis. Overall, 2799 Kcr sites were identified on 908 proteins with 14 conserved motifs. Bioinformatics analysis showed that Kcr was predominant on proteins found in cytoplasm, mitochondria and nucleus, and those involved in ribosome, proteasome, carbon metabolism and protein processing in endoplasmic reticulum. In total, 83 up-regulated and 12 down-regulated non-histone crotonylated sites were identified during spermiogenesis. These differentially expressed proteins were enriched in protein processing in endoplasmic reticulum pathway during formation of acrosome. In contrast, histone Kcr associated with mammalian spermatogenesis. These results provide foundational knowledge on the role of non-histone Kcr in spermiogenesis of E. sinensis. SIGNIFICANCE: Lysine crotonylation (Kcr) is a recently-identified post-translational modification, and histone Kcr was found to associate with mammalian spermatogenesis. However, crotonylation of non-histone proteins has not been reported in spermatogenesis regulation. Further, there is no information on crotonylation in crustaceans. This study was the first large-scale Kcr proteome characterization in crustaceans. A total of 2799 Kcr sites on 908 proteins with 14 conserved motifs were identified from Eriocheir sinensis testis. Of which, 83 up-regulated and 12 down-regulated non-histone crotonylated sites were identified during spermiogenesis. Our results provide the basic information for further functional validation of Kcr proteins and revealed new roles of Kcr in spermiogenesis of E. sinensis.
赖氨酸丁酰化（Kcr）是一种新发现的翻译后修饰类型。虽然 Kcr 在许多物种中都有报道，但在甲壳类动物中，人们对这一过程知之甚少。在这项研究中，采用泛抗赖氨酸丁酰化抗体富集和高分辨率液相色谱-质谱分析技术，对中华绒螯蟹（Eriocheir sinensis）精巢中的 Kcr 进行了表征。总体上，在 908 种蛋白质上鉴定出 2799 个 Kcr 位点，具有 14 个保守基序。生物信息学分析表明，Kcr 主要存在于细胞质、线粒体和细胞核中的蛋白质上，以及那些参与核糖体、蛋白酶体、碳代谢和内质网中蛋白质加工的蛋白质上。在精子发生过程中，共鉴定出 83 个上调和 12 个下调的非组蛋白丁酰化位点。这些差异表达的蛋白质在内质网中形成顶体的过程中被富集到蛋白加工途径中。相比之下，组蛋白 Kcr 与哺乳动物的精子发生有关。这些结果为非组蛋白 Kcr 在中华绒螯蟹精子发生中的作用提供了基础知识。
赖氨酸丁酰化（Kcr）是一种新发现的翻译后修饰，组蛋白 Kcr 与哺乳动物的精子发生有关。然而，非组蛋白蛋白的丁酰化在精子发生调控中尚未被报道。此外，甲壳类动物中也没有关于丁酰化的信息。本研究是甲壳类动物中首次大规模的 Kcr 蛋白质组学特征分析。从中华绒螯蟹精巢中鉴定出 908 种蛋白质上的 2799 个 Kcr 位点，具有 14 个保守基序。其中，在精子发生过程中鉴定出 83 个上调和 12 个下调的非组蛋白丁酰化位点。我们的研究结果为进一步对 Kcr 蛋白进行功能验证提供了基础信息，并揭示了 Kcr 在中华绒螯蟹精子发生中的新作用。
