Forschungsgruppe Peters

NANOMEDICINE TECHNOLOGIES

Scope of the group

The group is engaged in the field of nanomedicine which is understood (1) as the analysis, manipulation and application of the cellular nanomachinery to improve human health. In this context the group focusses on the development of devices composed of natural and artificial nanostructures. The hybrid devices shall be able to execute a variety of analytical and preparative tasks at efficiencies and speeds not previously achievable.

Our general approach for the development of hybrid devices comprises:

  • Analysis of cellular proteins or protein complexes in engineering terms.

  • Creation of core protein machines by recombinant methods and/or chemical synthesis.

  • Amalgamation of core machines with nanostructured solid-state substrates.


For the analysis of cellular protein complexes in vivo and in vitro we have developed fluorescence microscopic techniques such as photobleaching, optical single transporter recording and single molecule fluorescence microscopy. Among cellular protein complexes used as starting points for the creation of hybrid devices are the nuclear pore complex and the translocons of endoplasmic reticulum and mitochondria.

Photobleaching (FM, CFM, SCAMP)

The group has introduces, developed and applied a fluorescence microscopic technique (2) referred to as fluorescence microphotolysis (FM), fluorescence recovery after photobleaching (FRAP) or fluorescence photobleaching recovery (FPR). The method and various of its modifications such as CFM (3) and SCAMP (4) are widely used (5) for mobility measurements in living cells.

Optical Single Transporter Recording (OSTR)

The group has also developed (6-14) a nanotechnique referred to as Optical Single Transporter Recording (OSTR). OSTR is characterized by single transporter resolution, extreme sensitivity, transport substrate multiplexing, and parallel data acquisition. It can be combined with electrical single channel recording and single molecule fluorescence microscopy.

Single Molecule Fluorescence Microscopy (SMFM)

Recently, it has become possible to visualize single fluorescent molecules and to localize them at nanometer precission and track them at millisecond time resolution. The group has shown that Single Molecule Fluorescence Microscopy (SMFM) can be applied to three-dimensional systems and used to study the translational dynamics of single molecules in the cell nucleus (15-17).

Nuclear pore complex

The compartimentation of genetic material and protein synthetizing apparatus in the eukaryotic cell implies mechanisms for the selective and and regulated exchange of macromolecules between nucleus and cytoplasm. In that process the nuclear pore complex (NPC), a large structure made up of ~1000 protein, assumes a crucial role. The group has studied transport properties of the NPC by FM, OSTR and several other techniques (5,18).


REFERENCES

1. Peters, R. 2004. Perspectives of nanomedicine. Talk given at the International Symposium on Perspectives of Nanotechnology. Muenster, June 4-5, 2004. Manuscript can be obtained from the author.
2. Peters, R., J. Peters, K. H. Tews, and W. Bähr. 1974. A microfluorimetric study of translational diffusion in erythrocyte membranes. Biochimica et Biophysica Acta 367:282-294.
3. Peters, R., A. Brünger, and K. Schulten. 1981. Continous fluorescence microphotolysis: a sensitive method for study of diffusion processes in single cells. Proc. Natl. Acad. Sci. USA 78:962-966.
4. Wedekind, P., U. Kubitscheck, and R. Peters. 1994. Scanning microphotolysis: a new photobleaching technique based on fast intensity modulation of a scanned laser beam and confocal imaging. J. Microsc. 176:23-33.
5. Peters, R. 1986. Fluorescence microphotolysis to measure nucleocytoplasmic transport and intracellular mobility. Biochim. Biophys. Acta 864:305-359.
6. Peters, R., H. Sauer, J. Tschopp, and G. Fritzsch. 1990. Transients of perforin pore formation observed fluorescence microscopic single channel recording. EMBO J. 9:2447-2451.
7. Tschödrich-Rotter, M., U. Kubitscheck, G. Ugochukwu, T. Buckley, and R. Peters. 1996. Optical single channel analysis of the aerolysin pore in erythrocyte membranes. Biophys. J. 70:723-732.
8. Tschödrich-Rotter, M. and R. Peters. 1998. An optical method for recording the activity of single transporters in membrane patches. J. Microsc. 192:144-125.
9. Keminer, O. and R. Peters. 1999. Permeability of single nuclear pores. Biophys. J. 77:217-228.
10. Keminer, O., J. P. Siebrasse, K. Zerf, and R. Peters. 1999. Optical recording of signal-mediated protein transport through single nuclear pore complexes. Proc. Natl. Acad. Sci. USA 96:11842-11847.
11. Siebrasse, J. P., E. Coutavas, and R. Peters. 2002. Reconstitution of nuclear protein export in isolated nuclear envelopes. J. Cell Biol. 158:849-854.
12. Siebrasse, J. P. and R. Peters. 2002. Rapid translocation of p10/NTF2 through the nuclear pore of isolated nuclei and nuclear envelopes. EMBO reports 3:887-892.
13. Kiskin, N., J. P. Siebrasse, and R. Peters. 2003. Optical micro-well assay of membrane transport kinetics. Biophys. J. 2311-2322.
14. Peters, R. 2003. Optical Single Transporter Recording: Transport Kinetics in Microarrays of Membrane Patches. Annu. Rev. Biophys. Biomol. Struct. 32:47-67.
15. Kubitscheck, U., O. Kückmann, T. Kues, and R. Peters. 2000. Imaging of single GFP molecules in solution. Biophys. J. 78:2170-2179.
16. Kues, T., R. Peters, and U. Kubitscheck. 2001. Visualization and tracking of single protein molecules in the cell nucleus. Biophys. J. 80:2954-2967.
17. Kues, T., A. Dickmanns, R. Lührmann, R. Peters, and U. Kubitscheck. 2001. High intranuclear mobility and dynamic clustering of the splicing factor U1 snRNP observed by single particle tracking. Proc. Natl. Acad. Sci. USA 98:12021-12026.
18. Peters, R., E. Coutavas, and J. P. Siebrasse. 2002. Nuclear transport kinetics in microarrays of nuclear envelope patches. J. Struct. Biol. 140:268-278.


CONTACT

Prof. Dr. Reiner Peters
Institut für Medizinische Physik und Biophysik
Universität Münster
Robert-Koch-Straße 31
48149 Münster

Phone:+49 (0)251 835 69 33
Fax:+49 (0)251 835 51 21
In Organismen werden Stoffe und Informationen durch Kanäle ausgetauscht. Mit einer völlig neuen Methodik wird in der Medizinischen Physik die Funktionsweise einzelner Kanäle in Membranen untersucht.