Benndorf D, Loffhagen N, Babel W
Umweltforschungszentrum Leipzig-Halle GmbH, Sektion Umweltmikrobiologie, Germany.
Electrophoresis. 1999 Apr-May;20(4-5):781-9. doi: 10.1002/(SICI)1522-2683(19990101)20:4/5<781::AID-ELPS781>3.0.CO;2-A.
Cells of Acinetobacter calcoaceticus 69-V, a species able to metabolize a range of aliphatic hydrocarbons and alcohols, were confronted with ethanol, butanol, hexanol or heat shock during growth on acetate as sole source of carbon and energy. The primary alcohols and the heat shock led to the synthesis of new proteins or amplified expression of specific, common and general proteins, which were detected by silver staining after two-dimensional gel electrophoresis. Some of the alcohol-inducible proteins were identified as heat shock proteins by comparing protein patterns of alcohol-shocked cells with those of heat-shocked cells, and by N-terminal amino acid sequencing. DnaK was found to be amplified after all treatments, but GroEI only after heat shock and ethanol treatment. The N-terminal amino acid sequence of the protein, which was considerably amplified after alcohol treatment and heat shock, shows homology to HtpG (high temperature protein G). Some of the heat shock proteins induced by ethanol differ from those induced by butanol and hexanol, suggesting there are at least two different signals for the induction of some heat shock proteins by primary alcohols. This could be due to the different localization of ethanol, butanol and hexanol in the membrane, or because higher cytoplasmic concentrations of ethanol than of butanol or hexanol were applied in these tests in order to keep concentrations of the alcohols in the membrane roughly similar. Besides heat shock proteins, a group of proteins were observed which were only induced by butanol and hexanol, possibly indicating the existence of a further defense mechanism against high concentrations of hydrophobic substrates preventing protein denaturation and membrane damage.
醋酸钙不动杆菌69 - V能够代谢一系列脂肪族烃类和醇类，当该菌在以乙酸盐作为唯一碳源和能源的条件下生长时，使其分别接触乙醇、丁醇、己醇或进行热休克处理。在二维凝胶电泳后用银染法检测发现，伯醇和热休克处理导致了新蛋白质的合成或特定、常见及一般蛋白质表达量的增加。通过比较经醇类处理的细胞与经热休克处理的细胞的蛋白质图谱，并进行N端氨基酸测序，鉴定出一些醇类诱导蛋白为热休克蛋白。发现所有处理后DnaK蛋白的表达量均增加，但GroEI蛋白仅在热休克和乙醇处理后表达量增加。经醇类处理和热休克后大量增加的蛋白质的N端氨基酸序列与HtpG（高温蛋白G）具有同源性。乙醇诱导产生的一些热休克蛋白与丁醇和己醇诱导产生的不同，这表明伯醇诱导某些热休克蛋白至少存在两种不同的信号。这可能是由于乙醇、丁醇和己醇在膜中的定位不同，或者是因为在这些试验中使用的乙醇胞质浓度高于丁醇或己醇，以便使膜中醇类的浓度大致相似。除了热休克蛋白外，还观察到一组仅由丁醇和己醇诱导产生的蛋白质，这可能表明存在另一种针对高浓度疏水底物的防御机制，以防止蛋白质变性和膜损伤。
