Khandekar S S, Bettencourt B M, Kelley K C, Recny M A
Procept, Inc., Cambridge, Massachusetts 02139.
Protein Expr Purif. 1993 Dec;4(6):580-4. doi: 10.1006/prep.1993.1076.
GroEL, an Escherichia coli homolog of the heat shock protein 60 family of molecular chaperonins, has been implicated as a target of T cell-mediated immune responses in a broad spectrum of infections. In order to produce large quantities of native protein for raising and stimulating GroEL specific T cell lines, we have developed a simple and rapid two-step protocol for purifying native E. coli GroEL heat shock (or stress) protein which takes advantage of the inherent structural and functional properties of the protein. Based on a combination of gel exclusion chromatography, ATPase activity assay, isoelectric focusing, and circular dichroism analyses we conclude that our purification process yields native tetradecameric GroEL.
GroEL是分子伴侣热休克蛋白60家族的大肠杆菌同源物，在广泛的感染中被认为是T细胞介导的免疫反应的靶点。为了大量生产天然蛋白以培养和刺激GroEL特异性T细胞系，我们开发了一种简单快速的两步法方案，用于纯化天然大肠杆菌GroEL热休克（或应激）蛋白，该方案利用了该蛋白固有的结构和功能特性。基于凝胶排阻色谱、ATP酶活性测定、等电聚焦和圆二色性分析的组合，我们得出结论，我们的纯化过程产生了天然的十四聚体GroEL。
