Biochemistry Laboratory, ICAR-Central Inland Fisheries Research Institute, Barrackpore, Kolkata, 700120, India.
Appl Biochem Biotechnol. 2013 Oct;171(4):1011-21. doi: 10.1007/s12010-013-0384-y. Epub 2013 Aug 8.
Detection of species-specific sarcoplasmic peptides can be used as proteomic markers for fish food authentication and identification of species of origin in processed products. In the present study, proteomics technology was employed for differential characterization of sarcoplasmic peptides of two closely related fish species, Sperata seenghala and Sperata aor. Species-specific peptides were searched in white muscle extracts of the two species for identification of unique peptides that might aid in differentiation of the species, under two-dimensional gel electrophoresis platform. A total of 19 proteins were identified by combined matrix-assisted laser desorption ionization time-of-flight mass spectrometry and liquid chromatography-tandem mass spectrometry, of which nine and two proteins were found to be unique to S. seenghala and S. aor, respectively. One of the proteins, triosephosphate isomerase (TPI) was found to have three isoforms, out of which two were specific to S. aor, and one was specific to S. seenghala. All the three isoforms of TPI were present in the mixed samples of raw protein extracts of S. seenghala and S. aor, an observation that can be exploited to differentiate between the species and detection of deceptive practices of fraudulent substitution of commercially valuable fish species with inferior ones and differential characterization between closely related fish species.
肌浆肽的特异性检测可作为鱼类食品鉴伪和加工产品原产地鉴定的蛋白质组学标记。本研究采用蛋白质组学技术,对两种亲缘关系密切的鱼类,塞氏无须魮(Sperata seenghala)和印度无须魮(Sperata aor)的肌浆肽进行差异特征分析,以期在二维凝胶电泳平台下,找到有助于区分这两个物种的特异性肽。通过基质辅助激光解吸电离飞行时间质谱和液相色谱串联质谱联合鉴定,共鉴定出 19 种蛋白质,其中 9 种和 2 种蛋白质分别为 S. seenghala 和 S. aor 所特有。发现一种三磷酸甘油醛异构酶(TPI)有 3 种同工型,其中 2 种同工型为 S. aor 特有,1 种同工型为 S. seenghala 特有。所有 3 种同工型的 TPI 均存在于 S. seenghala 和 S. aor 的原始蛋白质混合提取物中,这一观察结果可用于区分这两个物种,检测商业价值高的鱼类被劣质鱼类欺诈性替代的欺诈行为,以及对亲缘关系密切的鱼类进行差异特征分析。