Kunugi S, Murakami Y, Ikeda K, Itoh N
Department of Polymer Science and Engineering, Kyoto Institute of Technology, Japan.
Int J Biol Macromol. 1992 Aug;14(4):210-4. doi: 10.1016/s0141-8130(05)80029-7.
High hydraulic pressure was used for denaturing proteins during chemical modifications. Bovine serum albumin and glucose oxidase were selected as the first targets of this unique technique and the ferrocene group was introduced into them, to obtain a macromolecular electron mediator and a self-electron-mediating oxidase, respectively. The result was compared with those obtained under non-denaturing conditions and under urea-denatured conditions. As for the number of ferrocene group linked to the protein, the pressure denaturation is superior to chemical denaturants, where ferrocenecarboxyaldehyde was used as the modifier. In both proteins the ferrocene group seemed to be introduced mainly inside the molecules with the pressure method, as the native conformation of the protein was restored when the high pressure was removed.
在化学修饰过程中，使用高液压来使蛋白质变性。选择牛血清白蛋白和葡萄糖氧化酶作为这种独特技术的首批目标，将二茂铁基团引入其中，分别获得一种大分子电子介体和一种自电子介体氧化酶。将结果与在非变性条件和尿素变性条件下获得的结果进行比较。至于与蛋白质相连的二茂铁基团的数量，当使用二茂铁甲醛作为改性剂时，压力变性优于化学变性剂。在这两种蛋白质中，通过压力法，二茂铁基团似乎主要引入到分子内部，因为当高压去除时蛋白质的天然构象得以恢复。
