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首页> 外文期刊>Bulletin of the Chemical Society of Japan >Design and Synthesis of Semi-Artificial Myoglobin Possessing DNA-Binding Peptides on Heme Propionates
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Design and Synthesis of Semi-Artificial Myoglobin Possessing DNA-Binding Peptides on Heme Propionates

机译：半人工肌红蛋白在血红素丙酸酯上具有DNA结合肽的设计与合成

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摘要

Peptides possessing the DNA-binding capability were successfully introduced into myoglobin by a cofactor-recon-stitution strategy.We have designed and synthesized an artificial iron-protoporphyrin IX(heme)-peptide conjugate,Heme(br)_2,which has covalently conjugated GCN4 basic region peptides at the heme propionate groups as an artificial DNA-binding site.The peptide Heme(br)2 was inserted into apomyoglobin to give a peptide-conjugated myoglobin,Mb(br)_2.The UV-vis and circular dichroism spectra of Mb(br)_2 were comparable to those of native myoglobin,suggesting that the heme environment and three-dimensional structure of Mb(br)_2 were almost identical to those of native myoglobin.The size exclusion chromatography indicated that the Mb(br)_2 existed as a monomeric form in an aqueous solution and interacted with its target DNA sequence [c-AMP responsive element(CRE)].The fluorescence study also revealed that the Mb(br)_2 bound CRE DNA effectively with 1/1 stoichiometry [affinity constant;8.4((+-)0.8)x 10~7 M~(-1)].Additionally,the CRE DNA-binding enhanced the peroxidase-like activity of Mb(br)_2,possibly due to the partial structural perturbation around the heme active-site.These results demonstrate that the chemical modification of prosthetic group with functional peptides can provide a new strategy for the design of semi-artificial proteins with engineered function.
机译：通过辅因子重组策略成功地将具有DNA结合能力的肽引入肌红蛋白中。我们设计并合成了人工共价结合GCN4的人工铁原卟啉IX（血红素）-肽结合物Heme（br）_2。将血红素丙酸酯基团上的碱性区域肽作为人工DNA结合位点。将血红素（br）2肽插入载脂蛋白中，得到肽结合的肌红蛋白Mb（br）_2.UV-vis和圆二色谱Mb（br）_2与天然肌红蛋白相当，表明Mb（br）_2的血红素环境和三维结构与天然肌红蛋白几乎相同。尺寸排阻色谱分析表明，Mb（br）_2以单体形式存在于水溶液中并与其靶DNA序列相互作用[c-AMP反应元件（CRE）]。荧光研究还表明，Mb（br）_2以1/1化学计量比有效结合CRE DNA [亲和力]康斯坦t; 8.4（（+-）0.8）x 10〜7 M〜（-1）]。另外，CRE DNA结合增强了Mb（br）_2的过氧化物酶样活性，可能是由于周围的部分结构扰动这些结果表明功能基团对修复基团的化学修饰可以为设计具有工程功能的半人工蛋白提供新的策略。

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