Nanotechnological Applications of Biomolecular Motor Systems. Stefan Diez Max-Planck-Institute of Molecular Cell Biology and Genetics Dresden

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1 Nanotechnological Applications of Biomolecular Motor Systems Stefan Diez Max-Planck-Institute of Molecular Cell Biology and Genetics Dresden

2 Max-Planck-Institute of Molecular Cell Biology and Genetics opened in scientists from 28 countries Biopolis Dresden

3 Generation and Manipulation of Nanostructures... Advantages: highly optimized processes small dimensions parallel processing possible molecular-specific interaction energy efficient potentially cheap... using Cellular Machines! Challenges: synthetic environment controlability reproducibility Kinesin + Microtubule

4 Kinesin walking on Microtubules Kinesin 70 nm step size: 8 nm speed: 0.8 µm/s (100 steps/s!!!) energy source: ATP hydrolysis Single Molecule Imaging

5 GFP-Kinesin Molecules Walking on Microtubules speed, run length, directionality, inhibition

6 Gliding Motility Assay to study Kinesin Gliding Assay control over motility 10 µm

7 Oriented Nanotransport on Artificial Substrates spatial guiding (topography, surface chemistry,...) temporal control (direction, speed,...) B cargo attachment (DNA, quantum dots...) A

8 Surface Topography Replica-Molding: Si Master PDMS Polyurethan (PU) channel width: 500 nm channel depth: 300 nm 500 nm PU-structure on glass directed movement changing between channels due to kinesin on the walls and rims

9 Surface Topography and Chemistry on Silicon I I I I I I I I I I I PEG C16 I I I I I I I I Kinesin I I I I I I I I xxxxxxxxxxxxxxxxxxxxxxxx Gold I I I I I I I I I I I directed movement of microtubules on Si-chip Nano Letters (5), 111, collaboration with group of Cees Dekker (TU Delft)

10 Rectification and Molecular Sorting rectification of motility molecular sorting Optimization of Rectification Geometries Nano Letters (5), 111, 2005.

11 Starting and Stopping Microtubule Motility... flowcell development with GeSiM GmbH... via ATP concentration volume of flow cell 6 µl MT x<5um_8mt MT 5um<x<10um_19MT MT x>10um_13mt Speed (µm/s) time (s) + ATP - ATP + ATP Microfluidics and Nanofluidics, in press, 2005.

12 Directing Motility in Hydrodynamic Flow straight flow zig-zag flow

13 Directing Motility by Electric and Magnetic Fields electrical fields / docking magnetic fields collaboration with TU Delft (C. Dekker) Nano Letters (5), 235, here: magnetic nanoparticles in solution planned: magnetic nanoparticles specifically attached to microtubules

14 Building Nano Wires and Networks with DNA Stretching DNA in a hydrodynamic flow gold SiO 2 SiO 2 gold DNA strand metallized with palladium bridging two gold electrodes (group of M. Mertig and W. Pompe, TU Dresden) Flow: 0.5 µl/s

15 Building Nano Wires and Networks with DNA Towards Nanoelectronics: gold Use biological machinery SiO 2 gold SiO 2 and evolutionary mechanisms of cells to build DNA strand metallized with palladium bridging two gold electrodes (group of M. Mertig and W. Pompe, TU Dresden) networks on the nanoscale.

16 Generation of DNA-based Nanocircuits Network Assembly Optimization Metallization gold contacts λ-dna microtubule restriction sites DNA binding to goldcontacts and microtubules stretching + transport of DNA by microtubules deposition of DNA on second gold contact creation periodic DNAnetwork by parallel processing structural selection by sequence-specific restriction enzymes deposition of metal complexes (Pt, Pd, Au, Ag)...or also carbonnanotubes, quantum dots...

17 Binding and Transport of DNA on motile Microtubules Nano Letters (3), 1251, Biotinylated Microtubules : - Biotin randomly incorporated - (9 tub / 3 Rhod-tub / 4 Biotin-tub) - labeled with Rhodamine λ-phage DNA ds : - 17µm long, 2nm in diameter - coiled molecules - biotinylated at both ends - labeled with YOYO

18 Pickup and stretching of DNA by motile MT 5 µm

19 Restriction of MT movement by stretched DNA... guiding of microtubules Nano Letters (3), 1251, 2003.

20 End-specific attachment of DNA to gold patterns gold gold

21 Other Projects Applications of Q-dots Biotemplated Nanostamping Protein Gradient Surfaces Nano Letters, in press, D Nanometry using FLIC Metallization of Microtubules PNAS, submitted, 2005.

22 Biomolecular Motors for Nanotechnology... use of cellular machines in synthetic environments is possible and attractive nano transport + structuring, molecular sorting Biomolecular Motors Nanotechnology surface chemistry + topography, quantum dots,...

23 Nanotechnology for Biophysics... application of nanotechnology to understand biophysics of motor proteins nano transport + structuring, molecular sorting Biomolecular Motors Nanotechnology surface chemistry + topography, quantum dots,...

24 Acknowledgements Collaborations: Jacob Kerssemakers, Bert Nitzsche, Felix Ruhnow (Single Molecule Techniques) Cordula Reuther, Cerasela Dinu, Leonid Ionov, Dimitar Stamov (Nanotec) Corina Bräuer (Technician) funded by the BMBF Nanotechnology Motor Proteins Biophysics: Joe Howard (MPI-CBG) Structures: T. Pompe (IPF Dresden) M. v. den Heuvel, C. Dekker (TU Delft) Flowcell: F. U. Gast, S. Howitz (GeSiM) DNA: J. Opitz, M. Mertig, W. Pompe (Max Bergmann Center of Biomaterials)