本研究旨在通过应用基于碘TMT的亚硝基蛋白质组学,用不同的处理方法(一氧化氮供体或一氧化氮合酶抑制剂)定量描述牛肉半膜(SM)中的S-亚硝基化。结果表明,在牛肉SM中368个蛋白中检测到2096个S-亚硝基化半胱氨酸位点。此外,筛选差异SNO修饰的蛋白质,其中一些涉及关键的生化途径,包括钙释放相关的蛋白质,能量代谢酶,肌原纤维,和细胞骨架蛋白。GO分析表明,差异蛋白位于广泛的细胞区室中,如细胞质,细胞器,和线粒体,为S-亚硝基化在死后肌肉中发挥广泛作用提供了先决条件。此外,KEGG分析验证了这些蛋白质参与了多种死后代谢过程的调节,尤其是糖酵解.最后,死后肌肉中S-亚硝基化水平的变化可能会影响关键肌肉蛋白的结构和功能,导致不同程度的肌肉代谢,最终影响牛肉品质。
This study aimed to quantitively profile the S-nitrosylation in beef semimembranosus (SM) with different treatments (nitric oxide donor or nitric oxide synthase inhibitor) by applying iodoTMT-based nitrosoproteomics. Results showed that 2096 S-nitrosylated cysteine sites in 368 proteins were detected in beef SM. Besides, differential SNO-modified proteins were screened, some of which were involved in crucial biochemical pathways, including calcium-releasing-related proteins, energy metabolic enzymes, myofibrils, and cytoskeletal proteins. GO analysis indicated that differential proteins were localized in a wide range of cellular compartments, such as cytoplasm, organelle, and mitochondrion, providing a prerequisite for S-nitrosylation exerting broad roles in post-mortem muscles. Furthermore, KEGG analysis validated that these proteins participated in the regulation of diverse post-mortem metabolic processes, especially glycolysis. To conclude, changes of S-nitrosylation levels in post-mortem muscles could impact the structure and function of crucial muscle proteins, which lead to different levels of muscle metabolism and ultimately affect beef quality.