MONOCYTE CHEMOATTRACTANT PROTEIN-1 AND ATHEROSCLEROSIS

描述（改编自研究者摘要）：单核细胞 趋化蛋白-1（MCP-1），一个相关的家族成员， 称为趋化因子的蛋白质是白细胞迁移的介质， 在健康和疾病中起着许多重要作用的激活， 包括炎症、动脉粥样硬化、癌症、伤口愈合和宿主 防御 MCP-1的产生和功能构成了一个目标， 临床干预来治疗多种此类疾病。 详细 了解MCP-1及其受体的结构和功能， 开发这种治疗策略是必要的。 MCP-1结构可以 基于其他趋化因子的结构建模，并且可能 由α螺旋和三链的球状结构域组成 β折叠 蛋白质的N-末端作为杆从蛋白质的末端延伸。 球状域 MCP-1通过结合许多 七跨膜受体家族成员的受体。 这些受体中的两种已被表征为MCP-1受体B （MCP-1 RB）（CC-CKR2 B）和巨细胞病毒产物开放阅读 帧US28（CMV-US28）。 CC-CKR2B对MCP-1具有特异性，而CMV-US28结合 单核细胞趋化蛋白-1和其他一些趋化因子。 长期目标是 这项研究旨在阐明MCP-1如何与这些受体结合， 信号进入目标细胞。 双管齐下的办法，包括协同 对受体和MCP-1的研究将用于建立理解 MCP-1/受体相互作用的研究 具体目标如下： 如下：1）进行MCP-1的诱变以发现MCP-1的功能重要性。 多肽的残基。 确定导致 通过在三个区域进行突变来实现MCP-1/受体相互作用： N-末端延伸杆，杆的基部区域和球状 结构域，并通过测量结合来确定生物学后果， 信号传导和趋化性。 确定MCP-1中负责 与糖胺聚糖结合。 2)进行氨基酸替换， MCP-I受体CC-CKR2B的进化保守残基，以发现 位点参与配体结合和信号转导。 3)确定 负责趋化因子受体特异性的序列元件， 产生两组嵌合受体：测量MCP 1与 RANTES与CC-CKR2B/CMV-US28嵌合体的结合和由结合引发的信号传导; 并测量MCP-1与IL 8的结合和信号转导参数 至CMV-US28/IL8RB嵌合体。 4)找到受体的区域， 通过发现MCP-1突变与MCP-1的特定残基相互作用， 差异性地影响与不同受体的结合， 信号转导，然后表征MCP-1的该子集的结合 突变体与嵌合受体的结合以及随之产生的信号传导。

DESCRIPTION (Adapted from Investigator's Abstract): Monocyte chemoattractant protein-1 (MCP-1), a member of the family of related proteins known as chemokines, is a mediator of leukocyte migration and activation that plays a number of significant roles in health and disease, including inflammation, atherosclerosis, cancer, wound healing and host defense. MCP-1 production and function constitute a target suitable for clinical intervention to treat a variety of such diseases. A detailed understanding of the structure and function of MCP-1 and its receptor is necessary to develop such therapeutic strategies. The MCP-1 structure can be modeled based on the structures of other chemokines and is likely to consist of a globular domain made up of an alpha helix and a three-strand beta sheet. The N-terminus of the protein extends as a rod from the globular domain. MCP-1 exerts its biological effects by binding to a number of receptors that are members of the seven transmembrane receptor family. Two of these receptors that have been characterized are the MCP-1 receptor B (MCP-1RB) (CC-CKR2B) and the product of the cytomegalovirus open reading frame US28 (CMV-US28). CC-CKR2B is specific for MCP-1 while CMV-US28 binds to MCP-1 plus a number of other chemokines. The long term goal of this research is to elucidate how MCP-1 binds to these receptors to transduce a signal into the target cell. A two-pronged approach, involving synergistic studies on both the receptors and MCP-1 will be used to build understanding of MCP-1/receptor interactions. The specific aims to be accomplished are as follows: 1)Perform mutagenesis of MCP-1 to find the functionally important residues of the polypeptide. Determine the residues responsible for MCP-1/receptor interactions by making mutations in three regions: the N-terminal extended rod, the region at the base of the rod, and the globular domain, and determine the biological consequences by measuring binding, signaling and chemotaxis. Identify the residues in MCP-1 responsible for binding to glycosaminoglycans. 2) Make amino acid substitutions in evolutionarily conserved residues of MCP-I receptors CC-CKR2B to find which sites are involved in ligand binding and signal transduction. 3) Determine the sequence elements responsible for chemokine receptor specificity by creating two sets of chimeric receptors: measuring binding of MCP1 versus RANTES to CC-CKR2B/CMV-US28 chimeras and signaling triggered by the binding; and measuring binding and signal transduction parameters of MCP-1 versus IL8 to CMV-US28/IL8RB chimeras. 4) Find regions of the receptors that directly interact with specific residues of MCP-I by finding MCP-I mutations that differentially affect binding to different receptors or the consequent signal transduction, then characterizing the binding of this subset of MCP-1 mutants to chimeric receptors and the consequent signaling.