BB 654

COURSE CANCELLED FOR WINTER 2008

Prerequisite: BB 452/552, BB 492/592, or consent of instructor.

Course content: Levels of protein structure, amino acid sequence determination, protein isolation and purification, chemical modification of proteins, spectroscopic approaches to protein structure, detection of protein-protein interactions, post-translational protein modification, and proteomics. In order to accommodate shifting interests, the class is consulted in determination of the emphasis that is placed on the various topics.

Text:
Required: Creighton, Proteins
Optional: Brandén & Tooze, Introduction to Protein Structure

Curricular purpose of the course: Presentation of experimental concepts and methods in understanding relationships of protein structure to function.

General Outline of Topics:

1. Review of Protein Structure. Review of protein secondary structure, models for protein folding, prediction of secondary structure from sequence, reversible denaturation.
2. Methods for protein isolation and purification. Extraction methods, selective precipitation techniques, chromatographic techniques (ion exchange, hydrophobic interaction, affinity).
3. Protein sequencing. Methods for amino acid analysis, preparation of fragments, review of the Edman degradatio, and mass spectrometry.
4. Post-translational Protein Modifications and Biological Control. Includes methods employed in the identification of phosphorylation sites with examples given demonstrating mass spectrometry, sequencing, and amino acid analysis.
5. Chemical Methods for the Study of Native Protein Structure. Active site labeling, covalent cross-linking, introduction of chromophores.
6. Spectroscopic Methods for the Study of Proteins in Solution. The related methods of ultraviolet absorption, circular dichroism, and fluorescence plus nuclear magnetic resonance.
7. Physical techniques to study protein-protein interactions in solution. Analytical sedimentation, fluorescence polarization, fluorescence resonance energy transfer, surface plasmon resonance.
8. Proteomics – overview.

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