Circular Dichroism Spectrometer for Protein/Peptide Structure/Function Studies

点击翻译按钮获取中文摘要

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.