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目的采用高分辨质谱方法研究蟾酥鲜品中的蛋白质。方法蟾酥鲜品蛋白经胰酶消化,利用纳升高效液相色谱-线性离子阱-静电场轨道阱(NanoLC-LTQ-Orbitrap)高分辨质谱技术对酶解多肽进行检测分析。Maxquant检索引擎鉴定蛋白[数据库为两栖纲蛋白序列(amphibia.fasta)]。结果鉴定了407种蛋白质和880种多肽序列。进一步采用基因本体论系统(gene ontology)中的PANTHER分析,对蟾酥鲜品的蛋白质组进行功能注释及分类。结论发现这些蛋白涉及多种信号通路,包括5-羟色胺(5-HT)受体介导通路,β肾上腺素能受体通路,血凝反应,钙信号通路和胆固醇合成与代谢等76种代谢路径。这为发现蟾酥鲜品中的蛋白质和其功能研究提供了实验依据。
Objective To study the protein content of fresh toad by high resolution mass spectrometry. Methods The fresh protein of Toad was digested by trypsin, and the digested protein was detected by NanoLC-LTQ-Orbitrap high performance liquid chromatography (HPLC). Maxquant search engine identifies proteins [database is amphibia.fasta]. As a result, 407 proteins and 880 polypeptide sequences were identified. The proteome of fresh toad was further annotated and classified using PANTHER analysis in gene ontology. Conclusions These proteins were found to be involved in a variety of signaling pathways including 76 metabolic pathways including serotonin (5-HT) receptor-mediated pathway, beta adrenergic receptor pathway, hemagglutination, calcium signaling and cholesterol synthesis and metabolism . This provides an experimental basis for the discovery of proteins and their functions in fresh toad products.