CHARACTERIZATION OF MAMMALIAN ADP-RIBOSYLTRANSFERASES

哺乳动物 ADP-核糖基转移酶的表征

• 批准号: 6109174
• 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
• 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/6109174
• 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

Mono-ADP-ribosylation is a post-translational modification of proteins in which the ADP-ribose moiety of NAD is transferred to proteins and is responsible for the toxicity of some bacterial toxins (e.g., cholera toxin, pertussis toxin). Similar to 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. The ART1 proteins were shown to be cell surface proteins, 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. In view of the expression of ARTs in tissues involved in immunoregulation, we examined whether ARTs might be expressed in pulmonary epithelial cells. Human airway epithelial cells were isolated by bronchoalveolar lavage and bronchial brushing. By in situ analysis and Northern blot, ART1 mRNA was identified in airway epithelial cells. As expected for GPI-anchored proteins, the localization of ART1 at the apical surface of ciliated epithelial cells was demonstrated by staining with polyclonal anti-ART1 antibody and confirmed by loss of this immunoreactivity after treatment with phosphatidylinositol-specific phospholipase C(PI-PLC), which selectively cleaves GPI-anchors and releases proteins from the plasma membrane. By in situ hybridization with specific ART3 and ART4 oligonucleotides, two additional members of the RT6 superfamily were also demonstrated in epithelial cells. In agreement with these findings, ART3 and ART4 mRNAs were identified by RT-PCR of poly(A)+RNA from human trachea. Interestingly, these proteins appeared to be preferentially localized to the airway epithelium. The localized expression of these members of the RT6 superfamily in human pulmonary epithelial cells may reflect a role in cell-cell signaling during immune responses within the airway.

单腺苷二磷酸核糖基化是一种翻译后 修饰蛋白质，其中NAD的ADP-核糖部分是 转移到蛋白质，并负责一些毒性 细菌毒素（例如，霍乱毒素、百日咳毒素）。类似于 霍乱毒素，一些哺乳动物ADP-核糖基转移酶 特别是利用精氨酸的胍基作为ADP-核糖， 接受者五种哺乳动物NAD：精氨酸ADP-核糖基转移酶 (ART)是从各种组织中克隆出来的ART 1蛋白是 显示是细胞表面蛋白，通过 糖基磷脂酰肌醇（GPI）锚。转移酶出现在 在哺乳动物组织中选择性表达。ART-1在 骨骼肌、心肌和淋巴样细胞，ART-2， 淋巴细胞，脾脏中的ART-4和睾丸中的ART-5。考虑到 在参与免疫调节的组织中， 检测了ARTs是否可能在肺上皮细胞中表达， 细胞人气道上皮细胞分离， 支气管肺泡灌洗和支气管刷拭。通过原位分析 北方杂交显示，ART 1 mRNA在气道上皮细胞中表达 细胞正如对GPI锚定蛋白所预期的， ART 1在纤毛上皮细胞的顶端表面， 通过用多克隆抗ART 1抗体染色证实， 证实了这种免疫反应性的损失后，处理与 磷脂酰肌醇特异性磷脂酶C（PI-PLC）， 选择性地切割GPI锚并从膜上释放蛋白质。 质膜通过与特异性ART 3和 ART 4寡核苷酸，RT 6的两个额外成员 在上皮细胞中也发现了超家族。一致 根据这些发现，ART 3和ART 4 mRNA被鉴定为 人气管poly（A）+RNA的RT-PCR。有趣的是，这些 蛋白质似乎优先定位于气道 上皮RT 6的这些成员的局部表达 人肺上皮细胞超家族可能反映了 在气道内的免疫反应期间的细胞-细胞信号传导。