Laboratory in Protein Purification and Enzymology

Catalog description

This course is intended to provide advanced Biochemistry & Molecular Biology students with in class instruction on various laboratory methods used to explore the physical & chemical properties of proteins and other topics relevant to enzymes such as protein purification techniques and enzyme kinetic assays. In addition students will have hands-on laboratory experience conducting experiments related to 1) enzyme kinetics and product inhibition pattern to determine the order of product release, 2) purification of proteins such as rabbit muscle aldolase using a variety of protein purification methods including salt precipitation, dialysis, and ion-exchange/affinity chromatography using phosphocellulose resin, 3) determination of specific activity and purity of a student purified protein, 4) determination of molecular weight of native enzymes using gel-filtration-HPLC and subunit molecular weight by SDS-PAGE, 5) determination of isoelectric point and identification of C-terminal amino acid of a purified protein, and 6) determination of optimal conditions for protein crystal growth. Student performance is evaluated through a variety of assessments, including lab reports organized in accordance to the biochemistry journal articles and through written examinations that assess the understanding of principles and methodology.