Circular Dichroism Spectrometer for Protein/Peptide Structure/Function Studies

圆二色谱(CD)是研究蛋白质和多肽在溶液中结构的重要工具。这项技术已被广泛用于在蛋白质中的特定氨基酸发生变化后探测蛋白质的结构。这项技术还被用于指导后续结构研究的蛋白质和多肽样品的制备。带有温度控制器的CD光谱偏振仪将被用来探测当重要氨基酸发生变化时蛋白质结构是否发生变化。正在研究的蛋白质是(1)金属-β-内酰胺酶(赋予细菌抗药性)，(2)3-酮酰基-ACP合成酶III(参与植物脂肪酸合成，并可能用于生产商业上感兴趣的油)，(3)乙醛基酶II，和(4)DEX1(参与减数分裂)。CD仪器还将用于指导核磁共振波谱研究样品的制备。这些研究涉及包埋在胶束中的跨膜蛋白、磷蛋白和植物去饱和酶/乙烯酶的特性。本科生、研究生、博士后助理和教职员工将使用圆二色光谱仪。本科生的具体使用包括由NSF资助的本科生研究体验(REU)计划支持的学生在生物化学实验室课程和物理化学实验室课程中的使用。该仪器将被安置在一个通用的系级仪器实验室中，迈阿密大学的所有研究人员都可以使用。圆二色光谱仪将为未来科学家的培训做出重大贡献，因为它为解决生物化学和生物物理学中的研究问题提供了重要的仪器。

Circular dichroism (CD) spectroscopy is an important tool for studying the structure of proteins and peptides in solution. The technique has been extensively used to probe the structure of proteins after specific amino acids in the proteins have been altered. This technique has also been used as a guide in the preparation of protein and peptide samples for subsequent structural studies. A CD spectropolarimeter with a temperature controller will be used to probe whether a change in the structure of proteins occurs when important amino acids are changed. The proteins under study are (1) metallo-beta-lactamase (which confers antibiotic resistance to bacteria), (2) 3-ketoacyl-ACP synthase III (which is involved in fatty acid synthesis in plants and can be potentially used to generate commercially-interesting oils), (3) glyoxylase II, and (4) DEX1 (which is involved in meiosis). The CD instrument will also be used to guide the preparation of samples to be studied with NMR spectroscopy. These studies involve the characterization of the membrane spanning proteins, phospholamban and plant desaturase/acetylenases, when embedded in micelles. Undergraduate students, graduate students, postdoctoral associates and faculty researchers will use the circular dichroism spectrometer. Specific use by undergraduates includes use by students supported under an NSF-funded Research Experiences for Undergraduates (REU) program, in a biochemistry laboratory course, and in a physical chemistry lab course. The instrument will be housed in a common-use, departmental instrumentation lab and will be accessible to all researchers at Miami University. The circular dichroism spectrometer will contribute significantly to the training of future scientists by providing access to important instrumentation for solving research problems in biochemistry and biophysics.