INDUCED CONFORMATIONS OF BIOLOGICALLY ACTIVE PEPTIDES

拟议研究的目的是研究诱导构象。 由多肽/脂和多肽/多肽相互作用产生。这将是 是通过研究诱导构象的扰动来实现的 反相高效液相色谱(RP-HPLC)在食品中的应用 结合圆二向色性(CD)。我们的一个基本要素 方法涉及完整系列的系统设计和综合 在起始亲本中只有一个变量的相关多肽 序列将在给定的时间被改变(即，单个氨基酸 省略、插入、替换等)。所有数据的结果 然后，研究的多肽将与它们的生物相关 活性，如抗菌活性和/或红细胞溶解。 导致生物活性的主要致病因素 多肽被认为是其诱导的二级结构 特定的行动地点。尽管多肽几乎总是用它们的 生物界面上的功能活动，特定的次级 它们在这样的界面上被诱导成的结构不能 很容易下定决心。核磁共振和X射线的技术改进 结晶学使这种构象研究得以进行 但这些技术非常耗时，而且仅限于 拥有所需物理和计算设施的实验室。 因此，我们将研究分子的相对诱导二级结构 反相高效液相色谱法测定特定的多肽系列 保留时间。通过这种方式，我们希望能够评估 反相高效液相色谱作为生物建模系统的潜在普遍用途 相关的水/脂界面。的简单性和可用性 反相高效液相色谱法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，合成 组合多肽库，由数千万个 肽，将用于研究蜂毒素与红色的相互作用部位 血细胞膜。这些方法预计将有一个 对多肽相互作用的理解产生重大影响 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.