THE POTENTIAL OF FLUORINATED SURFACTANTS IN MEMBRANE BIOCHEMISTRY
THE POTENTIAL OF FLUORINATED SURFACTANTS IN MEMBRANE BIOCHEMISTRY
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DOI：
10.1006/abio.1995.1003
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发表时间：
1995-01-01
影响因子：
2.9
通讯作者：
HOLZENBURG, A
中科院分区：
文献类型：
--
作者：
SHEPHERD, FH;HOLZENBURG, A
去污剂是膜生物化学中的重要试剂。由于所研究的每一种膜系统对去污剂的理化性能都有不同的要求，因此必须不断寻找新的生物化学去污剂。以全氟辛酸铵（APFO）为代表，对其作为“生物去污剂”的适宜性进行了研究。在洗涤剂浓度高达2% （w/v）时，它不会干扰Markwell对Lowry程序的修改。该洗涤剂的临界胶束浓度（cmc）值（0.013-0.0275 M）在一些具有生物学意义的缓冲液中进行了测定。结果表明，该去污剂可以通过透析去除，但速度较慢。这种缓慢的去除可能对重构/结晶研究特别有用。所列蛋白（液泡H+- atp酶膜区主要蛋白（16kda蛋白））的几种膜系统的增溶性研究；光系统II;马疱疹病毒（EHV）包膜蛋白)表明它是一种有效的增溶剂，可能在增溶性已经证明的情况下提高产量，并且在其他情况下，可以增溶以前难以增溶的蛋白质。研究了萃取-凝胶D树脂作为快速交换去污剂的方法，以最小的二次去污剂用量去除可溶性16kda蛋白中的APFO。(C) 1995学术出版社，Inc。
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. (C) 1995 Academic Press, Inc.