Single molecule methods are essential for gaining a thorough understanding of complex biological processes. This approach allows real time observation of signalling processes and of molecular machines at work. We use a variety of methods based on single molecule fluorescence and Förster Resonance Energy Transfer (FRET) to address dynamics on a wide timescale from less than a microsecond to several minutes. This is necessary for example for understanding how binding of small molecules (performed in less than a millisecond) can result in large conformational changes (taking up to seconds) and finally to regulation of protein expression levels in a cell.

The multidisciplinary nature of our research requires close collaboration with many colleagues in the fields of biology, biochemistry, medicine, biotechnology and physics.

Keywords:

Fluorescence, FRET, Single Molecule, Signalling, Molecular Machines, Hsp90, Conformational Dynamics

10 selected publications:

• Reliability and accuracy of single-molecule FRET studies for characterization of structural dynamics and distances in proteins.
  Agam G, Gebhardt C, Popara M, Mächtel R, Folz J, Ambrose B, Chamachi N, Chung S, Craggs T, de Boer M, Grohmann D, Ha T, Hartmann A, Hendrix J, Hirschfeld V, Hübner C, Hugel T, Kammerer D, Kang H, Kapanidis A, Krainer G, Kramm K, Lemke E, Lerner E, Margeat E, Martens K, Michaelis J, Mitra J, Moya Muñoz G, Quast R, Robb N, Sattler M, Schlierf M, Schneider J, Schröder T, Sefer A, Tan P, Thurn J, Tinnefeld P, van Noort J, Weiss S, Wendler N, Zijlstra N, Barth A, Seidel C, Lamb D & Cordes T (2023)
  Nat. Methods 20, 523-535. Doi: 10.1038/s41592-023-01807-0
• A blind benchmark of analysis tools to infer kinetic rate constants from single-molecule FRET trajectories.
  Götz M, Barth A, Bohr S, Börner R, Chen J, Cordes T, Erie D, Gebhardt C, Hadzic S, Hamilton L, Hatzakis S, Hugel T, Kisley L, Lamb C, de Lannoy C, Mahn C, Dunukara D, de Ridder D, Sanabria H, Schimpf J, Seidel C, Sigel R, Sletfjerding M, Thomsen J, Vollmar L, Wanninger S, Weninger K, Xu P, Schmid S (2022)
  Nat. Commun. 13(1) 5402. doi: 10.1038/s41467-022-33023-3.
• Angle-Dependent Strength of a Single Chemical Bond by Stereographic Force Spectroscopy.
  Cai W, Bullerjahn J T, Lallemang M, Kroy K, Balzer B N, Hugel T (2022)
  Chem. Sci., 13 (19), 5734–5740. Doi: 10.1039/D2SC01077A.
• Hierarchical Dynamics in Allostery Following ATP Hydrolysis Monitored by Single Molecule FRET Measurements and MD Simulations.
  Wolf S, Sohmen B, Hellenkamp B, Thurn J, Stock G, Hugel T (2021)
  Chem. Sci. 12: 3350-3359. doi: 10.1039/D0SC06134D. eLife 9: e57180. doi: 10.7554/eLife.57180.
• Controlling Protein Function by Fine-Tuning Conformational Flexibility.
  Schmid S, Hugel T (2020)
  eLife 9: e57180. doi: 10.7554/eLife.57180.
• Precision and Accuracy of Single-Molecule FRET Measurements - a Multi-Laboratory Benchmark Study.
  Hellenkamp B, Schmid S, Doroshenko O, Opanasyuk O, Kühnemuth R, Rezaei Adariani S, Ambrose B, Aznauryan M, Barth A, Birkedal V, Bowen M, Chen H, Cordes T, Eilert T, Fijen C, Gebhardt C, Götz M, Gouridis G, Gratton E, Ha T, Hao P, Hanke C, Hartmann A, Hendrix J, Hildebrandt L, Hirschfeld V, Hohlbein J, Hua B, Hübner C, Kallis E, Kapanidis A, Kim J, Krainer G, Lamb D, Lee N, Lemke E, Levesque B, Levitus M, McCann J, Naredi-Rainer N, Nettels D, Ngo T, Qiu R, Robb N, Röcker C, Sanabria H, Schlierf M, Schröder T, Schuler B, Seidel H, Streit L, Thurn J, Tinnefeld P, Tyagi S, Vandenberk N, Vera A, Weninger K, Wünsch B, Yanez-Orozco I, Michaelis J, Seidel C, Craggs T, Hugel T (2018)
  Nat. Methods 15: 669-676. doi: 10.1038/s41592-018-0085-0.
• Multidomain Structure and Correlated Dynamics Determined by Self-Consistent FRET Networks.
  Hellenkamp B, Wortmann P, Kandzia F, Zacharias M, Hugel T (2017)
  Nat. Methods 14: 174-180. doi: 10.1038/nmeth.4081.
• Four-Colour FRET Reveals Directionality in the Hsp90 Multicomponent Machinery.
  Ratzke C, Hellenkamp B, Hugel T (2014)
  Nat. Commun. 5: 4192. doi: 10.1038/ncomms5192.
• The large conformational changes of Hsp90 are only weakly coupled to ATP-hydrolysis.
  Mickler M, Hessling M, Ratzke C, Buchner J, Hugel T (2009).
  Nat. Struct. Mol. Biol. 16: 281-286. doi: 10.1038/nsmb.1557
• Single-Molecule Optomechanical Cycle.
  Hugel T, Holland N, Cattani A, Moroder L, Seitz M, Gaub H (2002)
  Science 296: 1103-1106. doi: 10.1126/science.1069856.