STRUCTURE DETERMINATION OF MITONEET, A POTENTIAL TARGET FOR DIABETES INHIBITION

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 在美国，肥胖症的增加与肥胖相关疾病的急剧增加相平行，最明显的是2型糖尿病。据预测，这种疾病在今后几十年内将达到流行病的程度。治疗II型糖尿病的首选药物是吡格列酮，这是一种噻唑烷二酮（TZD）衍生物，最初认为其通过激活核转录因子PPAR发挥作用。 最近，发现了吡格列酮的一个新的蛋白质靶点mitoNEET。 重组人mitoNEET的可溶形式在正交空间群中结晶，晶胞参数a = 46.81 A，B = 49.62 A，c = 59.01 A。结构测定为1.5 A分辨率。 从在对应于Fe的拐点、高能量远程和吸收峰的波长处收集的Fe-MAD数据集获得初始定相。 使用SSRL开发的自动MAD脚本处理数据。 该结构模型被细化到18.2%的R因子，并揭示了一种新的螺旋折叠，包含两个相距16 ½的2Fe-2S结合结构域，它们异常不稳定。蛋白质mitoNEET可能涉及Fe-S簇组装或储存，穿过细胞外膜的结合转移簇。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The rise in obesity in the United States parallels a dramatic increase in obesity-associated diseases, most notably type-2 diabetes. This disease is predicted to reach epidemic proportions in the next several decades. A drug of choice to treat type-II diabetes is pioglitazone, a thiazolidinedione (TZD) derivative originally thought to exert its effect through activation of the nuclear transcription factor PPAR. Recently, a novel protein target, mitoNEET, for pioglitazone was discovered. A soluble form of recombinant human mitoNEET was crystallized in an orthorhombic space group with unit-cell parameters a = 46.81 A, b = 49.62 A, c = 59.01 A. The structure was determined to 1.5 A resolution. Initial phasing was obtained from Fe-MAD datasets collected at wavelengths corresponding to the inflection, high energy remote, and absorption peak of Fe. Data was processed using automated MAD script developed at SSRL. The structural model was refined to an R-factor of 18.2 % and revealed a novel helical fold containing two 2Fe-2S binding domains 16 ¿ apart that are unusually labileThe protein mitoNEET may be involved with Fe-S cluster assembly or storage, binding transfer clusters that cross outer cell membranes.