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首页> 外文期刊>Molecular and Cellular Biology >Proteins binding to site C2 (muE3) in the immunoglobulin heavy-chain enhancer exist in multiple oligomeric forms.
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Proteins binding to site C2 (muE3) in the immunoglobulin heavy-chain enhancer exist in multiple oligomeric forms.

机译：与免疫球蛋白重链增强子中位点C2（muE3）结合的蛋白质以多种寡聚形式存在。

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We describe the purification to near homogeneity of proteins binding to site C2 (muE3) in the immunoglobulin heavy-chain enhancer. Proteins binding to this site produce four protein-DNA complexes which are distinguished by their mobility in gel retardation assays and their elution properties in an anion exchange column. DNA affinity-purified preparations of three chromatographically separated pools, containing different subsets of the four complexes, each contained three polypeptides of 42.5, 44, and 45 kilodaltons (kDa). UV crosslinking of protein to enhancer DNA demonstrated that site C2-binding activities in the three different pools bound DNA through proteins of similar sizes (about 45 kDa), even though the protein-DNA complexes formed by these binding activities were quite distinct. Gel exclusion chromatography and equilibrium binding analyses indicated that the distinct protein-DNA complexes were due to different oligomeric forms of the individual subunits and that a larger multimeric form bound with high affinity to the heavy-chain enhancer site C2, while a smaller species had a much lower affinity for heavy-chain enhancer sequences. Purified protein has been used to map high-affinity binding sites for site C2-binding proteins within an immunoglobulin heavy-chain promoter and at site KE3 in the kappa light-chain enhancer.
机译：我们描述了到免疫球蛋白重链增强剂中结合到位点C2（muE3）的蛋白质的接近同质的纯化。结合到该位点的蛋白质产生四种蛋白质-DNA复合物，这些复合物的特征在于它们在凝胶阻滞分析中的移动性以及它们在阴离子交换柱中的洗脱特性。 DNA亲和纯化的三个色谱分离库的制备物，包含四个复合物的不同子集，每个子​​集包含3个分别为42.5、44和45道尔顿（kDa）的多肽。蛋白质与增强剂DNA的UV交联表明，在三个不同池中的位点C2结合活性通过大小相似（约45 kDa）的蛋白质结合了DNA，即使由这些结合活性形成的蛋白质-DNA复合物非常不同。凝胶排阻色谱法和平衡结合分析表明，不同的蛋白质-DNA复合物是由于各个亚基的寡聚形式不同，较大的多聚体形式对重链增强子位点C2具有高亲和力，而较小的种类具有对重链增强子序列的亲和力要低得多。纯化的蛋白已被用于定位免疫球蛋白重链启动子内和Kappa轻链增强子中KE3位点C2结合蛋白的高亲和力结合位点。

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