Assigning mechanism to human polymorphisms via interacting residue identification

描述（由申请人提供）： 近年来大规模的人类单核苷酸多态性研究为揭示人类遗传变异与疾病的关系提供了丰富的信息。许多工具旨在通过使用物理和比较标准来预测SNP是否对蛋白质有害。另一方面，“变异可能对蛋白质无害，但会改变它与其相互作用伴侣相互作用的方式。因此，需要一种互补的方法来发现影响相互作用的突变。该提案绘制了一个系统的搜索，以寻找破坏蛋白质相互作用行为的遗传变异。首先，将采用相关突变分析来找到介导已知相互作用蛋白质对之间相互作用的氨基酸残基。随后将使用酵母双杂交方法（Y2 H）对假定的相互作用残基进行实验验证。然后，将确定位于这些残基的SNP，并最终使用Y2 H实验分析这些残基中的遗传变异如何破坏蛋白质-蛋白质相互作用。拟议的研究计划将发现人类蛋白质的微妙变化，这些变化影响它们如何与细胞中的其他蛋白质相互作用。这些发现的变异将为发现疾病的原因提供新的见解，并为我们理解遗传变异与细胞网络之间的关系增添新的维度。

DESCRIPTION (provided by applicant): The recent large-scale projects on human single nucleotide polymorphisms (SNP) provided a wealth of information to help uncover the relationship between human genetic variation and disease. A number of tools aims to predict if a SNP will be deleterious for a protein by using physical and comparative criteria. On the other hand,"amutation may not be deleterious for a protein, but change the way it interacts with its interaction partners. Hence, a complementary approach to find mutations that effect interactions are needed. This proposal maps out a systematic search for finding genetic variations that disrupt the interaction behavior of a protein. First, a correlated mutation analysis will be employed to find amino acid residues that mediate interaction between known interacting protein pairs. This will be followed by an experimental verification of the putative interacting residues using yeast two hybrid method (Y2H). Then, SNPs that are located at these residues will be identified and finally how the genetic variation in these residues disrupt protein-protein interactions will be experimentally analyzed using Y2H. The proposed research plan will find subtle variations in human proteins that influence how they interact with other proteins in the cell. These identified variations will offer insights for finding the causes of diseaes and also add a new dimension to our understanding of the relationship between genetic variation and cellular networks.