权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Identification of the functional domains in an orphan G protein-coupled receptor, GPR1, which acts as a coreceptor for the brain-derived cell tropism of HIV-1, HIV-2, and SIV

鉴定孤儿 G 蛋白偶联受体 GPR1 的功能域，该受体充当 HIV-1、HIV-2 和 SIV 脑源性细胞趋向性的辅助受体

基本信息

批准号：
12470068
负责人：
SHIMIZU Nobuaki
金额：
$ 3.2万
依托单位：
GUNMA UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470068/
关键词：
HIV-1 CORECEPTOR GPCR CELL TROPISM FUNCTIONAL DOMAIN GPCR HIV AIDS脳症 GPR1

项目摘要

【Aim】 Pericytes located around the brain blood vessels constitute the blood-brain barrier with endothelial cells. The infection of the cells with human immunodeficiency virus type-1 (HIV-1) is thought to be a causative factor to develop acquired immune deficiency syndrome (AIDS)-related encephalitis. An orphan G protein-coupled receptor, GPR1, acts as a coreceptor for HIV-1 strains which infect the brain vessels-derived pericytes. The aim of this study was to identify the important domain of GPR1 for its coreceptor activity. 【Methods】 (1) The amino-terminal domain (NTR) and three extracellular loops (ECLs) interact with HIV-1. Therefore, the DNA fragments of GPR1 coding four extracellular domains were prepared by polymerase chain reaction (PCR) and recombined with CCR5 which acts as a coreceptor for macrophage tropic HIV-1 strains. (2) Some nucleotide changes were introduced into GPR1 gene by PCR to make the amino acid deletion or substitution mutants of the NTR. (3) The chimeras betwe … More en GPR1 and CCR5 and the GPR1 mutants of the NTR were introduced into a glioma-derived cell line NP-2/CD4 which is strictly resistant to the infection with HIV-1, HIV type-2 (HIV-2), and simian immunodeficiency virus (SIV) even though the expression of CD4 is detected. Susceptibility of the established cells to various HIV-1, HIV-2, and SIV strains were detected. 【Results】 (1) All of ELC chimeras lost its coreceptor activities. (2) The NTR chimeras acted as the coreceptor. (3) The amino acid mutant of GPR1 in which the first tyrosine in the NTR was substituted with alanine lost the coreceptor function. On the other hand, the substitutions of the second, the third, and the fourth tyrosine with alanine had no effects on the function. (4) The deletion of the amino-terminal 11 amino acids of the NTR had no effects on the coreceptor activity of GPR1. 【Conclusion】 The NTR is important for the coreceptor function of GPR1. The region containing four tyrosine residues in the NTR is important for its coreceptor functions. These results are available to know the mechanisms of the HIV-1 infection to the brain vessels pericytes and applicable to the development of anti-HIV-1 drugs. Less

[Aim]位于脑血管周围的周细胞与内皮细胞共同构成血脑屏障。人类免疫缺陷病毒1型（HIV-1）感染细胞被认为是发展获得性免疫缺陷综合征（AIDS）相关脑炎的致病因素。孤儿G蛋白偶联受体GPR 1作为HIV-1毒株的辅助受体，HIV-1毒株感染脑血管源性周细胞。本研究的目的是确定其辅助受体活性的GPR 1的重要结构域。[方法]（1）HIV-1的氨基末端结构域（NTR）和三个胞外环（ECL）与HIV-1相互作用。因此，通过聚合酶链反应（PCR）制备了编码4个胞外结构域的GPR 1的DNA片段，并与作为嗜巨噬细胞的HIV-1株的辅助受体的CCR 5重组。(2)利用PCR技术对GPR 1基因进行核苷酸改变，制备NTR的氨基酸缺失或替换突变体。(3)我们之间的嵌合体 ...更多信息将GPR 1和CCR 5以及NTR的GPR 1突变体导入神经胶质瘤衍生的细胞系NP-2/CD 4中，该细胞系对HIV-1、HIV-2型（HIV-2）和猿猴免疫缺陷病毒（SIV）的感染具有严格抗性，尽管检测到CD 4的表达。建立的细胞对各种HIV-1，HIV-2和SIV株的敏感性进行了检测。[结果]（1）ELC嵌合体均丧失了辅助受体活性。(2)NTR嵌合体作为辅助受体。(3)GPR 1的氨基酸突变体，其中NTR中的第一个酪氨酸被丙氨酸取代，失去了辅助受体功能。另一方面，第二、第三和第四酪氨酸被丙氨酸取代对功能没有影响。(4)NTR氨基末端11个氨基酸的缺失对GPR 1的辅助受体活性没有影响。[结论] NTR对GPR 1的辅助受体功能有重要作用。NTR中含有四个酪氨酸残基的区域对其辅助受体功能很重要。这些结果有助于了解HIV-1感染脑血管周细胞的机制，并可用于抗HIV-1药物的开发。少