在本页:
Hunsicker-Wang实验室
生物化学 & 生物无机化学
概述: 研究 in the Hunsicker-Wang laboratory will focus on studying enzymes that utilize or bind metal ions, 所谓的金属离子. 有两个主要的兴趣领域:
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铁硫簇酶
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铁硫蛋白占所有金属蛋白的30%. These proteins utilize iron and sulfur atoms that are organized into clusters. 这些蛋白质经常参与电子转移反应. 具体地说, Rieske蛋白, 它是呼吸链复合体III的一部分, 包含一个[2Fe-2S]簇, which is ligated to the protein via 2 cysteine and 2 histidine residues. The reduction potential of this protein depends on the organism and the type of system that it was derived from. Previous studies have shown that the number of hydrogen bonds to the cluster, 溶剂可及性, and the type of charge residues near the cluster all affect the reduction potential. 对这种蛋白质的研究包括制造特定位点的突变, 净化, 结晶并解析突变酶的结构. 还对这些突变体的还原电位进行了评估. This protein is also be chemically modified with reagents that alter the properties of specific amino acids. This approach allows a greater variety of chemical properties to explore. Chemical modification also allows study of how individual amino acids contribute to the electron transport function within the protein.
Reactive oxygen species (ROS) are destructive and form from the reduction of molecular oxygen. One hypothesis is that a mismatch in the potential of Rieske蛋白 with its partners within Complex III leads to the production of ROS and may lead to neurodegenerative diseases. This hypothesis is starting to be explored in the Hunsicker-Wang lab.
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细胞色素氧化酶蛋白
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细胞色素氧化酶是呼吸系统的复合体. Within this protein, there are 4 metal sites, 2 heme-iron sites and 2 copper binding sites. One of the copper sites is the CuA center, found in subunit II of cytochrome oxidase. This center may also be involved in H+ translocation within cytochrome oxiase. 亚基II可以作为分离蛋白表达. The Hunsicker-Wang lab is exploring how the histidines in this protein may function to pump H+ using chemical modification and site-directed mutagenesis. 我们也在探索CuA蛋白是如何, Riesek蛋白, Azurin (a blue copper protein) and the Sco protein (a protein involved in the assembly of the CuA protein) are modified by endogenously produced molecules, 如4-羟基壬烯醛(HNE)和4-氧壬烯醛(ONE). These molecules are produced in membranes in the presence of reactive oxygen species such as peroxide. We are exploring how these molecules will react with amino acids of important metalloproteins involved in the respiration process.
maed实验室
生物化学 & 分子生物学
教授: 科瑞娜·梅德博士.D.
概述: 研究 in the maed实验室 centers on understanding the mechanisms involved in gene expression, 特别是前信使RNA剪接. 在真核生物, 最初, RNA transcribed from DNA may have intervening non-protein coding sequences, 或内含子. 为了准确地翻译蛋白质，必须去除这些序列. The removal of these introns must be precisely coordinated to avoid inaccuracies that can result in many diseases, 包括癌症和视网膜色素变性. 这个过程被称为前信使RNA剪接.
研究重点: A large macromolecular complex of RNA and proteins called the spliceosome facilitates splicing. The mechanism of pre-mRNA splicing involves large-scale rearrangements of protein-RNA complexes, which must be regulated to ensure both splicing timing and accuracy. Our research focuses on understanding these large-scale rearrangements within the spliceosome. The spliceosome is composed of five small nuclear ribonucleoprotein complexes (snRNPs). Dynamic rearrangements occur both within and between the snRNPs during the splicing cycle. These rearrangements are indicators that splicing is proceeding accurately. 不当拼接的后果是严重的. 在人类, 不当的剪接会导致一系列疾病, 包括色素性视网膜炎, 这是我们实验室的重点. Our research aims to dissect the molecular interactions that stimulate spliceosome assembly and activation. 具体地说, we are currently focused on how the interactions of splicing proteins Dib1, Prp6和Prp31有助于剪接体的组装. Dib, Prp6 and Prp31are essential for cell viability and splicing and are conserved from yeast to humans. 使用计算建模研究, 定点诱变和酵母生长试验, we have identified amino acids in each protein that are important for splicing. We are now trying to further characterize the interactions using biochemical and molecular biology techniques in order to understand how the interactions between proteins are help to maintain particular splicing complexes. 例如, a change to an amino acid in the Dib1 may cause the protein to not interact as well with Prp6 resulting in weakened interaction that stall spliceosome assembly. 整体, our work aims to build a molecular model for how these splicing proteins at the core of the splicing machinery help regulate spliceosome assembly directly or indirectly. 我们对剪接体的研究是跨学科的. 在我们的实验室, 我们使用各种生化, 分子生物学, and genetics techniques to dissect the importance of protein-nucleic acid and protein-protein interactions in the spliceosome. 学生也有学习生物化学的机会,包括 gel based binding assays for protein-RNA and protein-protein interactions, 结构研究使用圆二色性, 分子生物学, 使用蛋白质纯化, DNA克隆, and RNA transcription and purification and 3) genetics using Saccharomyces cerevisiae (Baker’s yeast) and mouse and human cell culture and 4) computational biochemistry, 通过我们与博士的合作项目. Kelvin Cheng in Physics in which students perform computational modeling studies on the spliceosome. These theoretical studies inform and parallel our experimental studies.