Fernandes P B, Nardi R V, Franklin S G
Institute for Cancer Research, Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111, USA.
Anal Biochem. 1978 Nov;91(1):101-14. doi: 10.1016/0003-2697(78)90820-5.
Currently available systems for resolving membrane proteins are based only on size and charge differences. Recently, it has been shown that Triton-urea-acetic acid gels which separate proteins on the basis of charge, size and hydrophobicity are capable of resolving proteins differing only by the substitution of a single neutral amino acid. We have applied this new method to the resolution of bacterial envelope proteins. Conditions for optimal resolution of different bacterial envelope proteins were determined by electrophoresis through transverse urea and Triton X-100 gradient gels. We have also correlated the components resolved in this system with those resolved by classical sodium dodecyl sulfate-gel electrophoresis by using two-dimensional slab gels combining the two systems. Furthermore, envelope protein fractions from different species and strains of bacteria were compared to identify specific proteins. This system appears to be a promising method for investigating envelope proteins which are due to missense mutations.
目前用于解析膜蛋白的系统仅基于大小和电荷差异。最近有研究表明，基于电荷、大小和疏水性分离蛋白质的Triton-尿素-乙酸凝胶能够解析仅因单个中性氨基酸取代而不同的蛋白质。我们已将这种新方法应用于细菌包膜蛋白的解析。通过横向尿素和Triton X-100梯度凝胶电泳确定了不同细菌包膜蛋白最佳解析的条件。我们还通过结合这两种系统的二维平板凝胶，将该系统中解析出的成分与经典十二烷基硫酸钠凝胶电泳解析出的成分进行了关联。此外，比较了来自不同细菌物种和菌株的包膜蛋白组分，以鉴定特定蛋白质。该系统似乎是研究由错义突变导致的包膜蛋白的一种有前景的方法。
