Fukuda R, Ishihama A, Saitoh T, Taketo M
Mol Gen Genet. 1977 Jul 20;154(2):135-44. doi: 10.1007/BF00330829.
The molecular structure of RNA polymerases from Escherichia coli, Salmonella typhimurium, Salmonella anatum,serratia marcescens, Aerobacter aerogens, Proteus mirabilis and Bacillus subtilis were compared based on:i) inhibition of the enzyme activity by treatment with antibodies against E. coli RNA polymerase subunits;ii) analysis of antibody precipitates by sodium ododecyl sulfatepolyacrylamide gel electrophoresis; and iii) analysis of antibody precipitates by urea-isoelectrofocusing followed by sodium dodecyl sulfate-slab gel electrophoresis in the second dimension. All the bacterial RNA polymerases examined cross-react equally with anti-E. COLI HOLOPOLYMERASE BUT EXHIbit different extents of cross-reaction with antibodies against individual subunits. Except for B. subtilis RNA polymerase, the molecular weight and isoelectric point of the enzyme subunits are close to those of E. coli polymerase. However, minor difference were found at least within the resolution of the techniques employed:S. anatum polymerase has sigma subunit larger than E. coli sigma subunit; P. mirabilis enzyme has sigma subunit larger in size and more acidic in charge, and alpha subunit smaller and more basic than corresponding E. coli subunits. The electrophoretic map of B. subtilis enzyme subunits is completely different from that of E. coli enzyme.
基于以下几点比较了来自大肠杆菌、鼠伤寒沙门氏菌、鸭沙门氏菌、粘质沙雷氏菌、产气气杆菌、奇异变形杆菌和枯草芽孢杆菌的RNA聚合酶的分子结构:i)用抗大肠杆菌RNA聚合酶亚基的抗体处理对酶活性的抑制作用;ii)通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析抗体沉淀物;iii)通过尿素-等电聚焦分析抗体沉淀物,然后在第二维进行十二烷基硫酸钠平板凝胶电泳。所有检测的细菌RNA聚合酶与抗大肠杆菌全酶的反应相同,但与抗单个亚基的抗体的交叉反应程度不同。除枯草芽孢杆菌RNA聚合酶外,酶亚基的分子量和等电点与大肠杆菌聚合酶的接近。然而,至少在所采用技术的分辨率范围内发现了微小差异:鸭沙门氏菌聚合酶的σ亚基比大肠杆菌的σ亚基大;奇异变形杆菌的酶的σ亚基更大且带更多负电荷,α亚基比相应的大肠杆菌亚基更小且带更多正电荷。枯草芽孢杆菌酶亚基的电泳图谱与大肠杆菌酶的完全不同。