The effects of an atpE ribosome-binding site mutation on the stoichiometry of the c subunit in the F1F0 ATPase of Escherichia coli.
The effects of an atpE ribosome-binding site mutation on the stoichiometry of the c subunit in the F1F0 ATPase of Escherichia coli.
复制标题
atpE 核糖体结合位点突变对大肠杆菌 F1F0 ATP 酶 c 亚基化学计量的影响。
DOI：
10.1006/abbi.1995.0063
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Brusilow,WS
中科院分区：
文献类型：
--
作者：
Schemidt,RA;Hsu,DK;Deckers-Hebestreit,G;Altendorf,K;Brusilow,WS
我们测试的假设，大肠杆菌F1 F0 ATP酶的F0扇区的C亚基的化学计量是依赖于atpE基因的表达水平。从携带编码F0亚基的质粒的细胞中纯化F0，所述F0亚基在编码c亚基的基因atpE之前具有或不具有核糖体结合位点突变。亚单位特异性抗体用于定量B和c亚单位的相对量。atpE表达的降低导致纯化的F0中c亚基的量显著降低。用免疫印迹法定量测定F1抗血清沉淀的F1 F0中B和c亚基的量也表明突变导致了c亚基化学计量的显著差异。从携带atpE核糖体结合位点突变的质粒合成的F1 F0中组装的c亚基的量比从野生型质粒合成的F1 F0中发现的量少2-5倍。因此，组装成F1 F0复合物的c亚基的化学计量似乎是可变的，这取决于atpE的表达。
We tested the hypothesis that the stoichiometry of the c subunit in the F0sector of the Escherichia coli F1F0ATPase is dependent upon the level of atpE gene expression. F0was purified from cells carrying plasmids encoding the F0subunits with and without a ribosome-binding site mutation preceding atpE, the gene which codes for the c subunit. Subunit-specific antibodies were used to quantitate the relative amounts of the b and c subunits. The decreased expression of atpE resulted in a significantly decreased amount of the c subunit in the purified F0. Immunoblot quantitation of the amounts of b and c subunits in F1F0precipitated by anti-F1antiserum also showed that the mutation produced significant differences in the stoichiometry of subunit c. The amount of c subunit assembled into the F1F0synthesized from a plasmid carrying the atpE ribosome binding site mutation was 2–5 times less than the amount found in the F1F0synthesized from a wild-type plasmid. Therefore, the stoichiometry of the c subunit assembled into the F1F0complex appears to be variable, depending on the expression of atpE.