Acquisition of Instrumentation for Peptide Synthesis and Purification

PI建议购买用于合成和纯化短肽的仪器。这一仪器将有助于推进PI的研究计划，包括模型α螺旋的光谱表征。长期以来，多肽的酰胺模式的红外光谱一直被用来估计二级结构含量和描述构象动力学。然而，使这些模式对骨架结构敏感的跃迁偶极耦合也使得这些模式成为蛋白质结构的全局标记，限制了它们作为特定的局部探针的应用。在这项研究中，引入到模型螺旋多肽序列中的同位素标记将被用来破坏过渡偶极耦合，并提供多肽结构和环境的局部报告。这些标记将被用来研究螺旋封顶和螺旋偶极对酰胺I频率的影响。此外，标记的残基将用于研究模型螺旋的折叠/去折叠过程，重点是特定残基的构象。关于局部相互作用对酰胺I频率影响的基本信息将是开发利用红外光谱作为探测蛋白质特定结构或动态性质的工具的方法学的基础。多肽合成仪器将使同位素标记多肽的现场制备成为可能，从而使这一研究项目的追求更具时间和成本效益。芒特霍利奥克学院是美国历史最悠久的女性持续文理学院，在培养女性从事科学研究方面有着悠久的历史。这一传统的一个关键组成部分是本科生和教职员工在研究项目上的合作。为了让学生为未来的研究事业做好最好的准备，本科生的研究项目必须尽可能多地让学生在技术和仪器方面有直接的实践经验。这种仪器将通过让学生接触到重要的现代生物化学技术来增强与PI的学生合作者的研究经验。

The PI proposes the acquisition of instrumentation for the synthesis and purification of short peptides. This instrumentation will help advance the research program of the PI involving spectroscopic characterization of model alpha helices. The infrared (IR) spectra of amide modes of peptides have long been used to estimate secondary structure content and describe conformational dynamics. However, the transition dipole coupling which makes these modes sensitive to backbone structure also causes these modes to be global markers about protein structure and limits their application as specific, local probes. In this study, isotopic labels introduced into the sequence of model helical peptides will be used to disrupt transition dipole coupling and to provide local reporters of peptide structure and environment. These labels will be used to investigate the effect of helix capping and helix dipoles on amide I frequency. In addition, the labeled residues will be used to investigate folding/unfolding processes of model helices focusing on the conformation of a particular residue. The basic information about the effect of local interactions on amide I frequencies will be the basis for developing methodology to use IR spectroscopy as a tool for probing of dissecting specific structural or dynamic properties of proteins. The peptide synthesis instrumentation will make the pursuit of this research project more time and cost efficient by making possible the preparation of isotopically labeled peptides on site. Mount Holyoke College, the nation's oldest continuing liberal arts college for women, has a long history of educating women for careers in scientific research. A key component of this tradition is the collaboration between undergraduate students and faculty on research projects. In order to best prepare students for future research careers, undergraduate research programs must give students as much direct, hands-on experience with the techniques and instrumentation as possible. This instrumentation will enhance the research experience of student collaborators with the PI by exposing them to important, modern techniques of biochemistry.