Protein Complexes: from Cells to Molecules

Module code: BS3070

Module co-ordinator: Dr Mohammed El-Mezgueldi

Practically all events in cells depend on the existence of protein complexes which interact with other components to bring about diverse cellular functions. Many molecular diseases arise from a failure in protein interactions and, in some cases, can be alleviated by specific drugs which act by blocking or modifying protein function.

In this module you will study functional protein complexes within cells, including how such complexes are identified and how their mechanism of action can be explored at the molecular level. We will use a range of in vitro and in vivo approaches to identify and characterise protein complexes, including fluorescence and NMR spectroscopy, crystallography, microscopy and yeast two-hybrid system.

The module will begin with an overview of protein complexes within cells and how they can be identified using mutational-, microscopy- and genomics-based methods. We will then cover high-throughput techniques (e.g. microarray) and other screening methods (e.g. yeast two-hybrid) to identify protein partners. We will discuss fluorescence methods in the context of measurements in living cells (microscopy) as well as analysis of interactions in vitro (spectroscopy). We will investigate structural methods (NMR and crystallography) which can define protein architecture and the changes in conformation that underlie the biological function of proteins. Bioinformatics is playing an increasing role in this area, as databases of protein structure continue to grow exponentially. Furthermore these databases can help identify structure-function relationships in novel proteins.

Topics covered

  • Evidence for the existence of protein complexes within cells
  • Techniques for identification of interacting partners: fluorescence spectroscopy and microscopy, yeast two-hybrid and phage display
  • Quantitative aspects of protein complexes: equilibrium binding, kinetics and thermodynamics of interactions
  • Techniques for studying protein structure: NMR and crystallography
  • Protein databases and the concept of protein docking

Learning

  • 32 one-hour lectures
  • 25 hours of practicals

Assessment

  • Exam, 3 hours (70%)
  • Data analysis (10%)
  • Literature analysis (10%)
  • Protein structure workshop (10%)