Guangdong Provincial Key Laboratory of Marine Biology, Shantou University, Shantou, 515063, China; Marine Biology Institute, Shantou University, Shantou, 515063, China; STU-UMT Joint Shellfish Research Laboratory, Shantou University, Shantou, 515063, China.
Guangdong Provincial Key Laboratory of Marine Biology, Shantou University, Shantou, 515063, China; Marine Biology Institute, Shantou University, Shantou, 515063, China; STU-UMT Joint Shellfish Research Laboratory, Shantou University, Shantou, 515063, China.
Dev Comp Immunol. 2023 Nov;148:104898. doi: 10.1016/j.dci.2023.104898. Epub 2023 Jul 31.
Lysine crotonylation (Kcr) is a newly discovered type of post-translational modification. Although Kcr has been reported in several species, its role in crustaceans remains largely unknown. In this study, Kcr in hemocytes of mud crab (Scylla paramamosain) was characterized using pan anti-crotonyllysine antibody enrichment and high-resolution liquid chromatogram-mass spectrometry analysis after SpTRAF6 or SpEcsit silencing. Altogether, 1,800 Kcr sites with six conserved motifs were identified from 512 proteins. Subcellular localization analysis showed that the identified Kcr proteins were mainly localized to the cytoplasm, nucleus, and mitochondria. The cellular components analysis showed that the 'chromosomal region' was enriched in the hemocytes of SpTRAF6-or SpEcsit-silenced mud crabs. The KEGG and PPI analyses showed that the identified Kcr proteins in the hemocytes SpTRAF6-or SpEcsit-silenced mud crabs were related to the 'protein processing in endoplasmic reticulum'; of which the marker of endoplasmic reticulum stress (Bip) was identified to be crotonylated. These datasets present the first comprehensive analysis of the crotonylome in mud crab hemocytes, providing invaluable insights into the regulatory functions of SpTRAF6 and SpEcsit in Kcr. Additionally, our findings shed light on the potential role of these proteins in activating marker proteins during endoplasmic reticulum stress in invertebrates.
赖氨酸丁酰化（Kcr）是一种新发现的翻译后修饰类型。尽管已经在几种物种中报道了 Kcr，但它在甲壳类动物中的作用在很大程度上仍然未知。在这项研究中，使用泛抗丁酰化赖氨酸抗体富集和 SpTRAF6 或 SpEcsit 沉默后高分辨率液相色谱-质谱分析，对泥蟹（Scylla paramamosain）血细胞中的 Kcr 进行了表征。总共从 512 种蛋白质中鉴定出 1800 个具有六个保守基序的 Kcr 位点。亚细胞定位分析表明，鉴定出的 Kcr 蛋白主要定位于细胞质、细胞核和线粒体。细胞成分分析表明，“染色体区域”在 SpTRAF6 或 SpEcsit 沉默的泥蟹血细胞中富集。KEGG 和 PPI 分析表明，SpTRAF6 或 SpEcsit 沉默的泥蟹血细胞中的鉴定出的 Kcr 蛋白与“内质网中的蛋白质加工”有关；其中内质网应激的标志物（Bip）被鉴定为丁酰化。这些数据集首次全面分析了泥蟹血细胞中的丁酰化组，为 SpTRAF6 和 SpEcsit 在 Kcr 中的调节功能提供了宝贵的见解。此外，我们的发现揭示了这些蛋白质在无脊椎动物内质网应激期间激活标记蛋白的潜在作用。
