ABZYME CATALYZED SITE-SPECIFIC ACYLATION OF PROTEINS

蛋白质的脂肪酸修饰是其中一个流行的主题 与信号有关的逆转录病毒蛋白和真核蛋白 转导。GAG多聚蛋白的N-肉豆蔻酰化是许多人所需要的 用于增殖的逆转录病毒和一些逆转录病毒癌蛋白 需要脂肪酸修饰才能转化细胞。因此，能够 调节蛋白质脂肪酸的酰化将是这项研究中有价值的工具 信号转导和逆转录病毒的生命周期。我们打算 催化抗体技术在蛋白质翻译后酰化反应中的应用 蛋白质。一种小分子膦多肽免疫小鼠的实验研究 半抗原应该导致产生一种特定于一种 所需的多肽序列由HAPEN预先确定。这样的abzyme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。我们的 这一领域的第一个目标将是确定abzyme是否针对 单一的三肽基磷酰胺可以区分半胱氨酸序列 以运动学上有用的方式从其他三肽中分离出来。这些实验 将使用三肽的组合固相库来完成，在 与一种新的图书馆筛选策略相结合。一次足够 选择性催化已经产生并分离出来，我们将启动 旨在酰化非肉豆蔻酰化形式的N-末端的实验 马克家族的人。这些实验构成了更广泛的 该项目的最终目标是应用催化抗体 技术对现场特定的翻译后修改 蛋白质。

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.