Hewick R M, Hunkapiller M W, Hood L E, Dreyer W J
J Biol Chem. 1981 Aug 10;256(15):7990-7.
A new miniaturized protein and peptide sequenator has been constructed which uses gas phase reagents at the coupling and cleavage steps of the Edman degradation. The sample is embedded in a matrix of Polybrene dried onto a porous glass fiber disc located in a small cartridge-style reaction cell. The protein or peptide, though not covalently attached to the support, is essentially immobile throughout the degradative cycle, since only relatively apolar, liquid phase solvents pass through the cell. This instrument can give useful sequence data on as little as 5 pmol or protein, can perform extended sequence runs (greater than 30 residues) on subnanomole quantities of proteins purified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and can sequence hydrophobic peptides to completion. The sequenator is characterized by a high repetitive yield during the degradation, low reagent consumption, low maintenance requirements, and a degradative cycle time of only 50 min using a complete double cleavage program.
一种新型的小型化蛋白质和肽测序仪已被构建出来，它在埃德曼降解的偶联和裂解步骤中使用气相试剂。样品被包埋在聚凝胺基质中，该基质干燥在位于小型盒式反应池中的多孔玻璃纤维盘上。蛋白质或肽虽然没有共价连接到支持物上，但在整个降解循环中基本保持不动，因为只有相对非极性的液相溶剂通过反应池。该仪器对低至5皮摩尔的蛋白质就能给出有用的序列数据，对于通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳纯化的亚纳摩尔量的蛋白质能够进行长序列测定（超过30个残基），并且能够对疏水肽进行完整测序。该测序仪的特点是降解过程中重复产率高、试剂消耗低、维护要求低，并且使用完整的双裂解程序时降解循环时间仅为50分钟。
