BIC-30803 Advanced Methods in Biochemical Research

Course

Credits 3.00

Language of instruction:

English

Assumed knowledge on:

BIC-20306 Cell Physiology and Genetics.

Continuation courses:

Thesis Biochemistry

Contents:

Note: This course has a maximum number of participants. The deadline for registration is one week earlier than usual. See 2014/2015 Academic Year (www.wageningenur.nl/en/Education-Programmes/Current-Students/Agenda-Calendar-Academic-Year.htm) -> Registration for Courses 2014-2015.

Advanced Methods in Biochemical Research is a multidisciplinary course in which different spectroscopic methods are used to unravel protein interactions and dynamics. In living cells protein interactions are pivotal in the regulation and control of a variety of processes in cells, such as growth and development. These processes function by signal transduction cascades, which are facilitated by e.g. conformational changes in proteins. Conformational changes of proteins and protein interactions can be monitored using various techniques of which Forster resonance energy transfer (FRET) is commonly applied.
In this course, various (spectroscopic) techniques are used to identify conformational changes of proteins. In vitro experiments, like steady state and time resolved spectroscopy, will be employed as well as single molecule experiments like fluorescence correlation spectroscopy. Protein complexes involved in regulatory or signal transduction pathways are isolated by affinity methods and the proteins in those complexes are identified by mass spectroscopy. In silico docking methods are applied to predict how proteins in a complex might interact.

Learning outcomes:

After successful completion of this course students are expected to:
- have knowledge on and insight into the application of advanced spectroscopic methods in studying cellular biochemistry;
- have experience with affinity purification, handling and characterization of proteins;
- have detailed knowledge about FRET methodology;
- be able to explain spectroscopic techniques such as steady state spectroscopy, time resolved fluorescence spectroscopy (anisotropy), fluorescence correlation spectroscopy and mass spectrometry;
- have insight in the application of sophisticated data analysis software for quantitative interpretation of protein interactions and dynamics.

Activities:

- performing experiments;
- analysing results;
- writing reports and making small Powerpoint presentations;
- participating in end-discussion.

Examination:

The practical course consists of two separate subjects; mass spectrometry (30%) and fluorescence spectroscopy (70%).
Marks will be given for:
- practical performance (15%);
- written reports (65%);
- oral presentation (20%).

Literature:

A manual that guides you through the experiments of this course is available on MyPortal.