RH PROTEIN AND THE RH BLOOD GROUP SYSTEM

RH 蛋白和 RH 血型系统

• 批准号: 6629154
• 负责人: CHEN-HAN HUANG
• 金额: $ 34.4万
• 依托单位: NEW YORK BLOOD CENTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-02-01 至 2005-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6629154
• 关键词: ABO(H) blood groups; antigen presentation; blood group antigens; blood groups; blood proteins; erythrocytes; erythroid stem cell; gene expression; gene mutation; gene targeting; genetically modified animals; glycoproteins; intracellular transport; laboratory mouse; laboratory rabbit; membrane transport proteins; molecular dynamics; mutant; peptides; phenotype; protein protein interaction; protein structure function; tissue /cell culture; transposon /insertion element

The RH family is composed of multiple polytopic membrane proteins present in a wide variety of species ranging from the most primitive unicellular slime mold to man. Two members, the variable RHCED polypeptides and the invariant RHAG glycoprotein, are restricted to red cells, defining the clinically important RH blood group system. These proteins have an important but unidentified function, as evidenced by changes in red cell morphology and physiology by products of their mutated genes. Based on our preliminary studies, we hypothesize that the RHAG homologs possess a conserved transport function in both erythroid and nonerythroid cells, while the RHCED series perform a related or modified function confined only to erythroid cells. We will examine this hypothesis by using model systems amenable to biochemical experimentation and genetic manipulation. Our long-term objectives are to define the molecular cellular requirement for RH expression, elucidate the function of the RH family proteins possibly as membrane transporters and unravel the individual and collective functional roles of RHAG/RHCED in red cells. To meet these goals, our specific aims are as follows. 1) To define the molecular cellular requirement for RH expression, we will use Rh-null mutations as a natural model and determine their mechanistic effects on cellular routing, membrane insertion and protein interaction. 2) To study the possible transport function, the slime mold ortholog RhgA will be employed as a simple model system. We will assess the function of RhgA by insertional gene inactivation and study the null mutants by expression rescue with wild type RhgA and human RH proteins. Candidate ligands will be sought and tested in transport assays. 3) To unravel the individual and collective functions of RHAG/RHCED, we will target-disrupt by homologous recombination the Rhced and Rhag orthologs in mice and determine the mutant red cell phenotypes. The proposed studies should lead to identification of the long-sought functional role of the RH family of proteins, and provide new insight into the structure/function relationships. The knowledge gained will also deepen our understanding of clinical problems associated with the RH blood group system, allowing the clinical service to explore new approaches for their management.

RH家族是由多个多位膜蛋白存在于各种各样的物种，从最原始的单细胞黏菌到man. Two成员，可变RHCED多肽和不变RHAG糖蛋白，仅限于红细胞，定义了临床上重要的RH血型系统。这些蛋白质具有重要但未鉴定的功能，如通过其突变基因的产物在红细胞形态和生理学上的变化所证明的。基于我们的初步研究，我们假设RHAG同源物在红系和非红系细胞中具有保守的转运功能，而RHCED系列仅限于红系细胞执行相关或修改的功能。我们将通过生物化学实验和遗传操作的模型系统来检验这一假设。我们的长期目标是确定RH表达的分子细胞要求，阐明RH家族蛋白可能作为膜转运蛋白的功能，并解开RHAG/RHCED在红细胞中的个体和集体功能作用。为了实现这些目标，我们的具体目标如下。1)为了定义RH表达的分子细胞要求，我们将使用Rh无效突变作为自然模型，并确定其对细胞路由，膜插入和蛋白质相互作用的机制影响。2)为了研究可能的转运功能，将采用黏菌直向同源物RhgA作为简单的模型系统。我们将通过插入基因失活评估RhgA的功能，并通过野生型RhgA和人RH蛋白的表达拯救研究无效突变体。将在转运试验中寻找和检测候选配体。3)为了阐明RHAG/RHCED的个体和集体功能，我们将通过同源重组靶向破坏小鼠中的Rhced和Rhag直系同源物，并确定突变的红细胞表型。拟议的研究应导致识别长期寻求的功能作用的RH家族蛋白质，并提供新的见解的结构/功能关系。所获得的知识也将加深我们对与RH血型系统相关的临床问题的理解，使临床服务能够探索新的管理方法。