ABZYME CATALYZED SITE-SPECIFIC ACYLATION OF PROTEINS

ABZYME 催化的蛋白质位点特异性酰化

• 批准号: 2608715
• 负责人: DARIN J GUSTIN
• 金额: $ 1.1万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-11-16 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2608715
• 关键词: abzyme; acylation; acyltransferase; haptens; laboratory mouse; posttranslational modifications; protein engineering

The modification of proteins with fatty acids is a prevalent theme among retro-viral proteins and eukaryotic proteins involved in signal transduction. N-myristoylation of the gag polyprotein is required by many retro-viruses for proliferation, and a number of retro-viral oncoproteins require fatty acid modifications transform cells. Thus, the ability to regulate protein fatty acid acylation would be a valuable tool in the study of signal transduction and the life cycle of retro-viruses. We intend to apply catalytic antibody technology to the post-translational acylation of proteins. Immunization of mice with a single small phosphono-peptido- hapten should result in the generation of actyltransferases specific to a desired peptide sequence predetermined by the hapen. Such abzymes could be used to block natural fatty acylation effectively interrupting signal transduction or interfering with the life-cycle of retro-viruses. Our first goal in this area will be to determine if abzymes elicited against a single tripeptido-phosphonamidate can distinguish the haptenic sequence from other tripeptides in a kinetically useful manner. These experiments will be done using a combinatorial solid phase library of tripeptides, in conjunction with a novel library screening strategy. Once sufficiently selective catalysis have been generated and isolated, we will initiate experiments aimed at acylating the N-terminus of the non-myristoylated form of the MARCKS. These experiments constitute the initial work in a broader project the ultimate goal of which is to apply catalytic antibody technology to the site specific post-translational modification of proteins.

用脂肪酸修饰蛋白质是一个普遍的主题， 参与信号转导的逆转录病毒蛋白和真核生物蛋白 转导 gag多聚蛋白的N-豆蔻酰化是许多 用于增殖的逆转录病毒，以及一些逆转录病毒癌蛋白 需要脂肪酸修饰来转化细胞。 因此， 调节蛋白质脂肪酸酰化将是一个有价值的研究工具 信号转导和逆转录病毒的生命周期。 我们打算 将催化抗体技术应用于 proteins. 用单个小膦酰基肽免疫小鼠， 半抗原应导致产生特异于 所需的肽序列由半抗原预先确定。 这种抗体酶可能是 用于阻断天然脂肪酰化，有效阻断信号 转导或干扰逆转录病毒的生命周期。 我们 这一领域的第一个目标是确定抗体酶是否引起抗 单个三肽-磷酰胺酸酯可以区分半抗原序列 从其他三肽中分离出来。 这些实验 将使用三肽的组合固相文库进行， 结合新的文库筛选策略。 一旦充分 选择性催化已经产生和分离，我们将启动 旨在酰化非豆蔻酰化形式的N-末端的实验 的马克。 这些实验构成了一个更广泛的 该项目的最终目标是将催化抗体 技术的位点特异性翻译后修饰 proteins.