CHARACTERIZATION OF MAMMALIAN ADP-RIBOSYLTRANSFERASES

哺乳动物 ADP-核糖基转移酶的表征

基本信息

批准号：
6432645
负责人：
Joel Moss
金额：
--
依托单位：
NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/6432645
关键词：
ADP ribosylation adenine phosphoribosyltransferase arginine bacterial toxins biochemical evolution enzyme activity enzyme mechanism enzyme structure human tissue laboratory mouse laboratory rabbit lymphoma molecular cloning protein sequence striated muscles

项目摘要

Mono-ADP-ribosylation is a post-translational modification of proteins in which the ADP-ribose moiety of NAD is transferred to protein and is responsible for the toxicity of some bacterial toxins (e.g., cholera toxin, pertussis toxin). Like cholera toxin, some mammalian ADP?ribosyltransferases specifically use the guanidino group of arginine as an ADP-ribose acceptor. Five mammalian NAD: arginine ADP-ribosyltransferases (ART) were cloned from various tissues. ART1 is a cell surface protein, linked through a glycosylphosphatidylinositol (GPI) anchor. The transferases appear to be selectively expressed in mammalian tissues. ART-1 is found in skeletal and cardiac muscle and lymphoid cells, ART-2 in lymphocytes, ART-4 in spleen and ART-5 in testis.The ADP-ribosyltransferases are characterized by a core catalytic domain that is conserved among the mammalian and bacterial enzymes. To define the requirements for ADP-ribosyltransferase activity, truncation mutants were made from the murine homologue of ART-1. This transferase catalyzes the ADP-ribosylation of arginine, and similar small guanidino compounds, as well as peptides and proteins containing arginine residues. In the absence of a guanidino acceptor for ADP-ribose, the transferase hydrolyzes NAD to ADP-ribose and nicotinamide, but at a much slower rate than arginine ADP-ribosylation. Removal of the signal sequences at the amino terminus, which is required for export into the endoplasmic reticulum, and at the carboxy terminus, which is necessary for addition of the GPI anchor, resulted in an active NAD:arginine ADP-ribosyltransferase. Further deletion of sequence from the amino terminus resulted in a significant loss of transferase activity, measured by ADP-ribosylation of both proteins and free arginine. In contrast, deletion of the amino terminal region of the protein increased NAD glycohydrolase activity, suggesting that the amino terminus has an inhibitory effect on abortive hydrolysis of NAD. In contrast, removal of amino acids at the carboxy terminus, which are closer to the putative catalytic site, resulted in a loss of both transferase and glycohydrolase activities.

单 ADP 核糖基化是蛋白质的翻译后修饰，其中 NAD 的 ADP 核糖部分被转移到蛋白质上，并导致某些细菌毒素（例如霍乱毒素、百日咳毒素）的毒性。与霍乱毒素一样，一些哺乳动物ADP核糖基转移酶专门使用精氨酸的胍基作为ADP核糖受体。从不同组织中克隆了五种哺乳动物 NAD：精氨酸 ADP-核糖基转移酶 (ART)。 ART1 是一种细胞表面蛋白，通过糖基磷脂酰肌醇 (GPI) 锚连接。转移酶似乎在哺乳动物组织中选择性表达。 ART-1 存在于骨骼、心肌和淋巴样细胞中，ART-2 存在于淋巴细胞中，ART-4 存在于脾脏中，ART-5 存在于睾丸中。ADP-核糖基转移酶的特点是具有在哺乳动物和哺乳动物中保守的核心催化结构域。细菌酶。为了确定 ADP-核糖基转移酶活性的要求，从 ART-1 的鼠同源物制备了截短突变体。该转移酶催化精氨酸和类似的小胍基化合物以及含有精氨酸残基的肽和蛋白质的 ADP-核糖基化。在不存在 ADP-核糖胍基受体的情况下，转移酶将 NAD 水解为 ADP-核糖和烟酰胺，但水解速度比精氨酸 ADP-核糖基化慢得多。去除氨基末端的信号序列（这是输出到内质网所需的）和羧基末端的信号序列（这是添加 GPI 锚所必需的），产生了活性 NAD:精氨酸 ADP-核糖基转移酶。通过蛋白质和游离精氨酸的 ADP 核糖基化测量，氨基末端序列的进一步缺失导致转移酶活性显着丧失。相反，蛋白质氨基末端区域的缺失增加了NAD糖水解酶的活性，表明氨基末端对NAD的无效水解具有抑制作用。相反，去除羧基末端的氨基酸（更接近假定的催化位点）会导致转移酶和糖水解酶活性丧失。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Characterization of NAD:arginine ADP-ribosyltransferases.

NAD：精氨酸 ADP-核糖基转移酶的表征。

DOI：
发表时间：
1999-03
期刊：
影响因子：
4.3
作者：
Moss, J;Balducci, E;Cavanaugh, E;Kim, H J;Konczalik, P;Lesma, E A;Okazaki, I J;Park, M;Shoemaker, M;Stevens, L A;Zolkiewska, A
通讯作者：
Zolkiewska, A

Modification of the ADP-ribosyltransferase and NAD glycohydrolase activities of a mammalian transferase (ADP-ribosyltransferase 5) by auto-ADP-ribosylation.

通过自动 ADP-核糖基化修饰哺乳动物转移酶（ADP-核糖基转移酶 5）的 ADP-核糖基转移酶和 NAD 糖水解酶活性。