兔肝金属硫蛋白亚型异构体的高效液相色谱分离与ESI-MS,MALDI-TOF-MS鉴定  

Separation of Rabbit Liver Metallothionein Sub-isoforms by RP-HPLC with MALDI-TOF-MS Detection

作　　者：赵锐[1] 纪建国[1] 袁洪生[1] 管华诗[2] 茹炳根[1] 

机构地区：[1]蛋白质组学研究室 [2]青岛海洋大学海洋药物与食品研究所,青岛266003

出　　处：《高等学校化学学报》2002年第6期1086-1090,共5页Chemical Journal of Chinese Universities

摘　　要：由于金属硫蛋白 (MT)基因的多态性 ,决定不同亚型的 MT异构体的存在 ,MT亚型异构体的结构是MT功能研究的基础 .通过离子交换柱可将 MT分成 MT-1和 MT-2两个异构体 ,用不同条件的反相高效液相色谱 (RP-HPLC)可将 MT-1和 MT-2分成不同的亚型异构体 ,并利用 MALDI-TOF MS和 LC-ESI-MS对比确定了它们的分子量 .结果表明 ,兔肝 MT在不同的 p H条件下分离得到不同分子量的亚型异构体 .在酸性条件下 ,MT-1可分为 2个主要亚型异构体 ,分子量分别为 61 49.0和 62 44.5 ,而 MT-2主要分为 3个亚型异构体 ,分子量分别为 61 49.0 ,62 44.0和 61 2 7.0 . MT-1和 MT-2有 2个亚型异构体分子量相同的异构体存在 .在酸性条件下 ,MT-1的 2个异构体及 MT-2分子量为 61 2 7的亚型异构体可稳定存在 .In order to study the sub isoforms of MT and get their exact primary structure, the mixture and the individuals MT 1 and MT 2 of rabbit liver MT were separated by different HPLC methods such as anion exchange and RP HPLC under different conditions. The two main MT isoforms MT 1 and MT 2 could be easily isolated from the mixture of MT by anion exchange HPLC. The isoforms of MTs were further isolated by C 18 RP HPLC under neutral and acidic conditions. All of the isolated isoforms and sub isoforms were analyzed by MALDI TOF MS. The separation of MT isoforms can be obtained by anion exchange chromatography with 50 mmol/L Na 2HPO 4 NaCl. The sub isoforms of rabbit liver MT could be isolated by C 18 RP HPLC. The rabbit liver MT 1 has 3 main sub isoforms and MT 2 has one sub isoform. The experiment verified that under different conditions the sub isoforms of rabbit liver MT has a different apparent molecular mass. This experiment predicted that there would exist a stable peptide structure of MT 2 or its fragment. The primary structure of this peptide component was also studied.

关 键 词：高效液相色谱 分离 鉴定 金属硫蛋白 亚型异构体 电喷雾离子化质谱 基质辅助激光解析飞行时间质谱 

分 类 号：Q51[生物学—生物化学] Q503