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Single Molecule Investigations of Protein Dynamics

Prof. Don Lamb, Chemistry Department, Ludwig-Maximilians-University Munich (LMU)

 

With current technology, it is possible to perform experiments on individual molecules. In my group, we are developing these advanced fluorescence technologies to extract the maximum amount of information possible from single molecule experiments and are using this technology to determine how proteins function.

One of the main methods we use is Fluorescence Resonance Energy Transfer or FRET. FRET is a phenomenon where energy is transferred between fluorescence molecules via dipole-dipole interactions. The rate of energy transfer depends on the distance between them and is sensitive to distances in the range of 2-10 nm, making it an ideal ruler to investigate the dynamics of molecules and molecular interactions. Typically, FRET is performed with two fluorophores. However, with additional fluorophores, more distances can be investigated simultaneously and new types of information gathered in the experiment. For example, whether motions in different domains of a protein are correlated or independent.

The goal of this PhD project is to establish three and four color FRET and use this technique to investigate the conformational dynamics of proteins and protein complexes. The project focuses on the use of burst analysis with multiparameter fluorescence detection and pulsed-interleaved excitation (Kudryavtsev, 2012, ChemPhysChem, 13:1060) for multicolor single molecule FRET experiments. The experience gained during the project includes development of the instrumentation, advancement of the analysis software and the preparation, labeling and measurement of biological samples.

Candidates must be interested in interdisciplinary research, optical microscopy, quantitative analysis and biology. A degree in physics, physical chemistry, engineering, biochemistry or a related subject is required. Experience in optics or fluorescence microscopy and cell biology is beneficial but not necessary.

 

More information about the group:
Prof. Lamb's website