Nucleotide binding by a 24-residue peptide from the RecA protein of Escherichia coli.
Nucleotide binding by a 24-residue peptide from the RecA protein of Escherichia coli.
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来自大肠杆菌 RecA 蛋白的 24 个残基肽与核苷酸结合。
DOI：
10.1073/pnas.83.24.9289
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发表时间：
1986
影响因子：
11.1
通讯作者：
McEntee,K
中科院分区：
文献类型：
--
作者：
Knight,KL;McEntee,K
我们最近证明了两种ATP类似物亲和标记物，8-叠氮腺苷5 '-三磷酸（N3 ATP）和5'-对氟磺酰基苯甲酰基腺苷（5 'FSBA），在位于跨越残基257-280的24-残基胰蛋白酶肽（T-31）内的特定酪氨酸残基（Tyr-264）处共价修饰大肠杆菌的RecA蛋白。在这里，我们表明，N3 ATP有效地修饰纯化的肽T-31，并表明，相互作用是特定的以下标准：肽T-31的光标记是饱和的N3 ATP浓度，光标记是竞争性的ATP和腺苷，但不与腺嘌呤，UTP，或TTP;和其他肽衍生自RecA蛋白质是穷人的光标记的底物，除了一个片段，表现出非特异性相互作用的光亲和类似物。N3 ATP修饰的T-31的分析表明，光标记附着到一个以上的网站内的肽。这些数据表明，肽T-31含有一些位点的腺嘌呤和核糖部分的ATP结合时，它的RecA蛋白的接触。
We have recently demonstrated that two ATP analog affinity labels, 8-azidoadenosine 5'-triphosphate (N3ATP) and 5'-p-fluorosulfonylbenzoyladenosine (5'FSBA), covalently modify RecA protein of Escherichia coli at a specific tyrosine residue (Tyr-264) located within a 24-residue tryptic peptide (T-31) spanning residues 257-280. Here we show that N3ATP efficiently modifies purified peptide T-31 and show that the interaction is specific by the following criteria: photolabeling of peptide T-31 is saturable with respect to the N3ATP concentration; photolabeling is competitive with ATP and adenosine but not with adenine, UTP, or TTP; and other peptides derived from RecA protein were poor substrates for photolabeling except for one fragment that showed a nonspecific interaction with the photoaffinity analog. Analysis of N3ATP-modified T-31 shows that the photolabel attaches to more than one site within the peptide. These data argue that peptide T-31 contains some sites of contact for adenine and ribose moieties of ATP when it is bound to RecA protein.