JNY-2中抗肿瘤活性相关蛋白成分的鉴定

Identification of antitumor protein components in JNY-2

目的:鉴定海洋中药魁蚶(JNY-2)中发挥抗肿瘤作用的蛋白成分.方法:运用JNY-2转录组测序reads,de novo组装unigenes,构建JNY-2转录组数据库;通过液质联用技术联合转录组数据,对分离得到的JNY-2活性蛋白组分JNY-2-1中发挥抗肿瘤作用的物质基础进行分析鉴定.结果:通过对海洋中药JNY-2进行转录组测序,共得到142 280 214条reads,de novo组装后得到132 025个unigene,构建了JNY-2转录组ORF数据库;对JNY-2抗肿瘤活性蛋白组分JNY-2-1进行LC-MS/MS分析,产生了574 120个MS/MS二级质谱图谱,与JNY-2 (ORF)数据库比对吻合且因假发现率(FDR)≤1%的有9 101个MS/MS光谱;鉴定得到特征肽段2 381个,最终在活性组分JNY-2-1中鉴定了516个蛋白成分,包含调控生物进程的功能性蛋白和细胞构成蛋白,但无毒素类蛋白成分.结论:从JNY-2抗肿瘤活性蛋白组分JNY-2-1鉴定得到516个蛋白成分,为活性新蛋白的发现及海洋中药JNY-2抗肿瘤物质基础研究提供了重要的科学资料.

Objective: To identify the antitumor protein components of Scapharca broughtonii (JNY-2), a marine traditional Chinese medicine. Methods: Unigenes was assembled by sequencing the transcriptome of Scapharca broughtonii by reads and de novo, and the database of transcriptome was constructed. The antitumor substance basis of the active protein components JNY-2-1, isolated from JNY-2, was identified by using liquid-mass spectrometry combined with transcriptome data analysis. Results: According to the transcriptome sequencing of S. broughtonii , 142 280 214 reads were obtained, 132 025 unigenes was obtained via de novo assembly, and an ORF database of S. broughtonii transcriptome was constructed. The antitumor active protein components JNY-2-1 was analyzed by LC-MS/MS, and 574 120 MS/MS spectra were produced, which matched the ORF database of the S. broughtonii with 9 101 MS/MS spectra of a FDR not more than 1%. There were 2 381 characteristic peptides identified, and 516 protein components were identified in JNY-2-1, including functional proteins regulating biological processes, cell constituent proteins, and no toxin proteins. Conclusion: 516 protein components were identified from JNY-2-1. This study provides the scientific information for the discovery of new active proteins and the study of the antitumor substance basis of S. broughtonii .