CHEM 642 - Protein Chemistry Techniques

Units: 3
Techniques in protein chemistry. Methods for protein quantification, separation, identification and structure determination. Emphasis on modern methods such as liquid chromatography, electrophoresis, isoelectric focusing, expression vectors, immunological probing, mass spectrometry and computer-based analysis. Lecture Units: 1; Lab Units: 2.

Prerequisites: CHEM 340 or CHEM 441, both with grade C- or better.
Possible Instructional Methods: Entirely On-ground.
Grading: A-F grading only.
Student Learning Outcomes - Upon successful completion of this course students will be able to:

List several types of differential solubility and column chromatography techniques used in protein purification and explain the mechanism for each.
Apply the separation techniques of ammonium sulfate precipitation and group-specific column chromatography to purification of an enzyme and quantify the enzyme activity in international units.
Contrast the theoretical basis for HPLC (High Performance Liquid Chromatography) with that for FPLC (Fast Protein Liquid Chromatography) and utilize these techniques to separate protease-digested protein fragments and protein mixtures, respectively.
Appraise the advantages of using recombinant DNA techniques to express normal or hybrid proteins prior to purification and isolate a hybrid protein by affinity chromatography.
Relate the principles underlying non-denaturing and denaturing polyacrylamide gel electrophoresis (PAGE) and isoelectric focusing.
Utilize SDS-PAGE and immunoblot detection to analyze protein induction from an expressionvector, purification of a hybrid protein, and reductive separation of IgG subunits.
Apply the techniques of IEF and SDS-PAGE to the 2D separation of a complex protein mixture and compare this technique with protein microarray technology.
Explain the principles of mass spectrometry (MS) as applied to the analysis of peptides derived from proteins identified in proteomics experiments.
Make use of online databases (UniProtKB/SWISS-PROT, RCSB-PDB) to identify protein sequences and 3D structures, and deduct structural information for an uncharacterized protein sequence using programs such as ExPASY, Predict Protein and SWISS-MODEL.