CARHOHYDRATE MOIETIES OF GP120

GP120 的碳水化合物部分

• 批准号: 3144086
• 负责人: KHUSHI L MATTA
• 金额: $ 23.13万
• 依托单位: ROSWELL PARK CANCER INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-09-01 至 1994-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3144086
• 关键词: AIDS; HIV envelope protein gp120; affinity chromatography; bovine serum albumin; carbohydrate structure; chemical synthesis; enzyme inhibitors; enzyme linked immunosorbent assay; glycoproteins; glycosidases; glycosylation; goats; human immunodeficiency virus; human subject; laboratory rabbit; mannose; mannosidase; mass spectrometry; neutralizing antibody; nuclear magnetic resonance spectroscopy; serology /serodiagnosis; tissue /cell culture

The envelope glycoprotein gp120 of the human immunodeficiency virus (HIV) is a high glycosylated molecule. It has been suggested that gp120 plays a vital role in viral attachment and the initiation of infection through interaction of CD4 glycoprotein of T-lymphocytes. This program centers at the chemical synthesis of oligosaccharide structures which are being reported as part of the carbohydrate moiety of gp120 and their further use as immunogens. Attention will be focussed on the synthesis of glycosides that possess an aglycon which can serve as a bridge for attachment to high molecular weight substances such as bovine serum albumin. Complex structures containing mannose is one of our major interests in this program. We plan to investigate the use of various derivatives of 4- pentenyl-alpha-D-mannopyranoside as glycosylating agents. The structures of our synthetic compounds will be established by n.m.r. studies and mass spectroscopy. Our synthetic antigens will be further utilized to raise their antibodies. The availability of antigen moiety linked to a solid support (Sepharose) will facilitate purification of the antibodies raised against the corresponding antigens. Specificity of the purified antibodies will be established with a battery of our well-defined carbohydrate structures. Upon purification the antibodies produced to the synthetic obligosaccharides will be tested for their ability to block gp120-CD4 binding and gp120-CD4 mediated cell fusion and for their ability to neutralize virus growth. Our synthetic saccharides linked to BSA will be used to develop saccharide ELISA methodology for the detection of anti- carbohydrate antibodies in the sera of AIDS patients. Thus, in the program we also plan to screen AIDS patients' sera for well-defined oligosaccharide specific antibodies. In another aspect of this program, we will emphasize our studies toward the chemical synthesis of various potential inhibitors of glycosidases, such as glucosidases and mannosidases, involved in the processing and biosynthesis of glycoprotein including gp120. The effect of glycosylation inhibitors on gp120 maturation will be assessed in a variety of transfected cell lines and in virus-infected cells. Our potential inhibitors may prove to be chemotherapeutic agents for AIDS.

人类免疫缺陷病毒包膜糖蛋白gp120 是一种高度糖基化的分子。已有研究表明gp120在 在病毒附着和通过以下途径启动感染中起重要作用 T淋巴细胞CD4糖蛋白的相互作用。该计划的中心是 低聚糖结构的化学合成 报告为gp120碳水化合物部分的一部分及其进一步的用途 作为免疫原。重点关注糖苷类化合物的合成。 它拥有一种配基，可以作为连接HIGH的桥梁 分子量物质，如牛血清白蛋白。 含有甘露糖的复杂结构是我们在这方面的主要兴趣之一 程序。我们计划调查不同的衍生产品的用途4- 戊烯-α-D-甘露糖苷作为糖基化试剂。这些结构 我们的合成化合物的一部分将由n.m.r建立。研究和质量 光谱学。我们的合成抗原将被进一步利用来提高 他们的抗体。与固体相关联的抗原部分的可用性 载体(琼脂糖凝胶)将促进所提抗体的纯化 对抗相应的抗原。纯化抗体的特异性 将由我们定义明确的碳水化合物电池建立起来 结构。在纯化后，产生的抗体合成的 将测试必需糖阻止gp120-cd4的能力。 结合和gp120-CD4介导的细胞融合及其能力 中和病毒生长。我们与牛血清白蛋白相连的合成糖将是 用于建立检测抗-HBs的糖类ELISA法 艾滋病患者血清中的碳水化合物抗体。因此，在节目中， 我们还计划筛选艾滋病患者血清中明确定义的寡糖 特定的抗体。 在这个计划的另一个方面，我们将强调我们对 化学合成多种潜在的糖苷酶抑制剂，如 参与加工和生物合成的葡萄糖苷酶和甘露糖苷酶 包括gp120在内的糖蛋白。糖基化抑制剂对血管内皮生长因子的影响 Gp120的成熟度将在各种转基因细胞系中进行评估。 以及在感染病毒的细胞中。我们的潜在抑制因素可能会被证明是 艾滋病的化疗药物。