INDUCED CONFORMATIONS OF BIOLOGICALLY ACTIVE PEPTIDES

本研究的目的是研究诱导构象 由肽/脂质和肽/肽相互作用产生。 这将 通过研究诱导构象的扰动来完成 使用反相高效液相色谱法（RP-HPLC）， 圆二色性（CD） 在我们的生活中， 方法涉及系统的设计和综合的完整系列 在起始亲本中只有一个变量 序列将在给定时间改变（即，单个氨基酸 省略、插入、替换等）.所有的结果 研究的肽将与它们的生物学特性相关联。 活性，如抗微生物活性和/或红细胞裂解。 主要的致病因素负责的生物活性， 肽被认为是其诱导的二级结构， 具体的行动地点。 虽然肽几乎总是发挥其 在生物界面的功能活动，特定的次级 它们在这种界面处被诱导成的结构不能被 容易确定。 核磁共振和X射线技术的改进 晶体学使这类构象研究得以进行， 但是这些技术非常耗时，并且仅限于 拥有所需物理和计算设施的实验室。 因此，我们将研究相对诱导的二级结构， 通过RP-HPLC变化测定的特定肽系列 保留时间。 通过这种方式，我们希望能够评估 RP-HPLC作为生物学建模系统的潜在普遍用途 相关的水/脂质界面。 的简单性和可用性， RP-HPLC将使大多数实验室能够利用我们的 当前和未来的结果。 作为一种补充方法， 组合肽库，由数千万个 肽，将用于研究蜂毒素的网站的相互作用与红色 血细胞膜 这些方法预计将有一个 对理解肽相互作用的重大影响， 免疫学、分子生物学和微生物学领域。

The objective of the proposed research is to study induced conformations resulting from peptide/lipid and peptide/peptide interactions. This will be accomplished by studying the perturbations of induced conformations using reversed-phase high-performance liquid chromatography (RP-HPLC) in conjunction with circular dichroism (CD). An essential element in our approach involves the systematic design and synthesis of complete series of related peptides in which only one variable in the starting parent sequence will be altered at a given time (i.e. , a single amino acid omitted, inserted, substituted, etc.) . The results for all of the peptides studied will be then correlated with their biological activities, such as antimicrobial activity, and/or red blood cell lysis. The dominant causative factor responsible for the biological activity of peptides is believed to be their induced secondary structures at their specific sites of action. Although peptides virtually always exert their functional activity at biological interfaces, the specific secondary structures into which they are induced at such interfaces cannot be readily determined. Technological improvements in NMR and X-ray crystallography enable conformational studies of this kind to be carried out, but these technologies are time-consuming and limited to those laboratories with the required physical and computing facilities. Therefore, we will study the relative induced secondary structures of specific peptide series as determined by variation of their RP-HPLC retention times. In this way, we expect to be able to assess the potential general usefulness of RP-HPLC as a system modeling biologically relevant aqueous/lipid interfaces. The simplicity and availability of RP-HPLC will enable the majority of laboratories to take advantage of our current and future results. As a complementary approach, synthetic combinatorial peptide libraries, comprised of tens of millions of peptides, will be used to study melittin's sites of interaction with red blood cell membranes. These approaches are expected to have a significant impact on the understanding of peptide interactions in the fields of immunology, molecular biology, and microbiology.