Vibrational Stark Effect in Human Aldose Reductase

人醛糖还原酶(HALR2)在高血糖条件下将葡萄糖还原为山梨醇， 被认为是糖尿病长期影响健康的原因，包括白内障和肾病。一键 寻找不与结构相似的蛋白质人醛结合的hALR2的强抑制剂 还原酶(HALR1)，是测定抑制物结合部位的静电场。在中测量这些字段 这种复杂的系统将使用hALR2结合的振动斯塔克效应(VSE)光谱来完成 使用含有氰基官能团的抑制剂，该氰基官能团在 光谱中没有蛋白质吸收的区域。首先，一系列的抑制分子将被 氰基的位置在蛋白质的疏水口袋周围移动的合成 活动站点。其次，将获得蛋白质-抑制物复合体的晶体结构。最后，氨基酸 活性部位附近的残基将发生突变，以改变探针周围的局部静电场。VSE 然后将对结合了活性部位的抑制剂的蛋白质突变体进行光谱分析。结果将 与hALR2中当前静电场计算模型的预测进行比较。

Human aldose reductase (hALR2) reduces glucose to sorbitol under hyperglycemic conditions, and is thought to be a cause of long-term health effects of diabetes including cataracts and nephropathy. One key to finding strong inhibitors of hALR2 that do not also bind to a structurally similar protein, human aldehyde reductase (hALR1), is to measure the electrostatic field of the inhibitor binding site. Measuring these fields in such a complicated system will be done using vibrational Stark effect (VSE) spectroscopy of hALR2 bound with inhibitors which contain a cyano functional group, which has a vibrational absorption frequency in a region of the spectrum that is free of protein absorption. First, a series of inhibitor molecules will be synthesized in which the location of the cyano group is moved around the hydrophobic pocket of the protein active site. Second, a crystal structure of the protein-inhibitor complex will be obtained. Finally, amino acid residues near the active site will be mutated to alter the local electrostatic field surrounding the probe. VSE spectroscopy will then be conducted on protein mutants with inhibitor bound to the active site. Results will be compared to predictions from current computational models of electrostatic fields in hALR2.