Shepherd F H, Holzenburg A
Department of Biochemistry and Molecular Biology, University of Leeds, United Kingdom.
Anal Biochem. 1995 Jan 1;224(1):21-7. doi: 10.1006/abio.1995.1003.
Detergents are important reagents in membrane biochemistry. Since each membrane system studied places different demands on the detergent in terms of desirous physicochemical properties, detergents new to biochemistry must continuously be sought. Ammonium perfluorooctanoate (APFO) was investigated, as representative of fluorinated surfactants, in terms of its suitability as a "biological detergent." It did not interfere with the Markwell modification of the Lowry procedure at detergent concentrations of up to 2% (w/v). Critical micellization concentration (cmc) values (0.013-0.0275 M) for this detergent were determined in a number of buffers of biological interest. It was demonstrated that the detergent can be removed by dialysis, albeit slowly. This slow removal may be particularly useful for reconstitution/crystallization studies. Solubilization studies on several membrane systems containing the proteins listed (the major protein of the membrane sector of the vacuolar H(+)-ATPase (16 kDa protein); photosystem II; equine herpes virus (EHV) envelope proteins) indicate that it is a potent solubilizing agent, likely to enhance the yield in cases where solubilization has already been demonstrated, and, in other cases, to solubilize proteins formerly recalcitrant to solubilization. The removal of APFO from solubilized 16-kDa protein by means of Extracti-Gel D resin as a means of exchanging detergents quickly and with a minimum requirement for second detergent was investigated.
去污剂是膜生物化学中的重要试剂。由于所研究的每个膜系统对去污剂的理想物理化学性质都有不同要求，因此必须不断寻找生物化学领域新型的去污剂。作为含氟表面活性剂的代表，全氟辛酸铵（APFO）被作为一种“生物去污剂”来研究其适用性。在去污剂浓度高达2%（w/v）时，它不会干扰洛瑞法的马克韦尔改良法。在多种具有生物学意义的缓冲液中测定了这种去污剂的临界胶束浓度（cmc）值（0.013 - 0.0275 M）。结果表明，该去污剂可以通过透析去除，尽管速度较慢。这种缓慢去除对于重组/结晶研究可能特别有用。对几种含有所列蛋白质的膜系统（液泡H(+) - ATP酶膜区的主要蛋白质（16 kDa蛋白质）；光系统II；马疱疹病毒（EHV）包膜蛋白）进行的增溶研究表明，它是一种有效的增溶剂，在已经证明可以增溶的情况下可能提高产量，而在其他情况下，能够增溶以前难以增溶的蛋白质。研究了通过Extracti - Gel D树脂从溶解的16 kDa蛋白质中去除APFO，以此作为一种快速交换去污剂且对第二种去污剂需求最小的方法。
