SYNTHETIC FLAVO HEMO PROTEINS

我们已经成功地设计和建造了从头合成 具有光激活蛋白内电子的黄血黄蛋白 调职。我们使用我们传统的四螺旋捆绑框架来 将溴黄素共价连接到内部的半胱氨酸上， 束的几何中心。在相同的捆绑包中，内饰 组氨酸A与添加的亚铁血红素形成双-希氏配位 朝向束状物内部的任一端。我们已经证明了 蓝色黄素吸收带的照明产生了一个激发 一种能从一种方便的电子中提取一对电子的状态 含氮碱基供体(如EDTA)可产生还原的黄素 自发地将电子转移到附近连接的双组氨酸 亚铁血红素。我们的目的是使用RLBL纳秒染料激光器来 捕获激发的黄素态的光谱，随后 减少了黄素，并提供了时间的初步描述 蛋白质内电子从还原型黄素转移到 亚铁血红素。很有可能会有一种异质性 与黄素/血红素分布相对应的时间进程 氧化还原肽不同构象的距离。我们是 黄素激发态猝灭的竞争研究 向亚铁血红素的能量转移和亚麻素之间的电子转移 还有亚铁血红素。我们正在运送人工合成的黄血球蛋白 乙二胺四乙酸乙二酯溶液通过电池比较区域的光谱 而且没有蓝色泵浦脉冲。我们很容易通过以下方式再生我们的材料 将溶液暴露在分子氧中，这会迅速氧化 血红素成分。

We have succeeded in designing and constructing de novo synthetic flavo-hemo-proteins that exhibit light activated intraprotein electron transfer. We use our traditional four-helix bundle framework to covalently link a bromoflavin to an interior cysteine very close to the geometric center of the bundle. In the same bundle, interior histidines a positioned to form a bis-His ligation of added heme toward either end of the bundle interior. We have shown that illumination of the blue flavin absorption band creates an excited state which can extract a pair of electrons from a convenient nitrogenous base donor (such as EDTA) to create a reduced flavin that spontaneously transfers electrons to nearby bis-histidine ligated heme. It is our intent to use the RLBL nanosecond dye laser to capture the spectra of the excited flavin state, the subsequent reduced flavin, and to provide an initial description of the time course of intraprotein electron transfer from the reduced flavin to the heme. It is quite possible that there will be a heterogeneity of the time course corresponding to a distribution of flavin/heme distances for different conformations of the redox peptide. We are investigating a competition between flavin excited state quenching by energy transfer to the heme and electron transfer between the flavin and the heme. We are flowing the synthetic flavo-hemo-protein plus EDTA solution through a cell to compare the spectra of regions with and without the blue pump pulse. We easily regenerate our material by exposing the solution to molecular oxygen, which rapidly oxidizes the heme component.