Tag Archives: Philipp C. Nickels

A DNA origami-based nanoscopic force clamp

Nanoclamp made of DNA strands. Illustration: Christoph Hohmann

Nanoclamp made of DNA strands. Illustration: Christoph Hohmann

An Oct. 21, 2016 news item on ScienceDaily announces a new nanotool,

Physicists at Ludwig-Maximilians-Universitat (LMU) in Munich have developed a novel nanotool that provides a facile means of characterizing the mechanical properties of biomolecules.

An Oct. 21, 2016 Ludwig-Maximilians-Universitat (LMU) press release (also on EurekAlert), which originated the news item, explains the work in more detail (Note: A link has been removed),

Faced with the thousands of proteins and genes found in virtually every cell in the body, biologists want to know how they all work exactly: How do they interact to carry out their specific functions and how do they respond and adapt to perturbations? One of the crucial factors in all of these processes is the question of how biomolecules react to the minuscule forces that operate at the molecular level. LMU physicists led by Professor Tim Liedl, in collaboration with researchers at the Technical University in Braunschweig and at Regensburg University, have come up with a method that allows them to exert a constant force on a single macromolecule with dimensions of a few nanometers, and to observe the molecule’s response. The researchers can this way test whether or not a protein or a gene is capable of functioning normally when its structure is deformed by forces of the magnitude expected in the interior of cells. This new method of force spectroscopy uses self-assembled nanoscopic power gauges, requires no macroscopic tools and can analyze large numbers of molecules in parallel, which speeds up the process of data acquisition enormously.

With their new approach, the researchers have overcome two fundamental limitations of the most commonly used force spectroscopy instruments. In the case of force microscopy and methodologies based on optical or magnetic tweezers, the molecules under investigation are always directly connected to a macroscopic transducer. They require precise control of the position of an object – a sphere or a sharp metal tip on the order of a micrometer in size – that exerts a force on molecules that are anchored to that object. This strategy is technically extremely demanding and the data obtained is often noisy. Furthermore, these procedures can only be used to probe molecules one at a time. The new method dispenses with all these restrictions. “The structures we use operate completely autonomously“, explains Philipp Nickels, a member of Tim Liedl’s research group. “And we can use them to study countless numbers of molecules simultaneously.”

A feather-light touch

The members of the Munich group, which is affiliated with the Cluster of Excellence NIM (Nanosystems Initiative Munich), are acknowledged masters of “DNA origami”. This methodology exploits the base-pairing properties of DNA for the construction of nanostructures from strands that fold up and pair locally in a manner determined by their nucleotide sequences. In the present case, the DNA sequences are programmed to interact with each other in such a way that the final structure is a molecular clamp that can be programmed to exert a defined force on a test molecule. To this end, a single-stranded DNA that contains a specific sequence capable of recruiting the molecule of interest spans from one arm of the clamp to the other. The applied force can then be tuned by changing the length of the single strand base by base. “That is equivalent to stretching a spring ever so-o-o slightly,” says Nickels. Indeed, by this means it is possible to apply extremely tiny forces between 1 and 15 pN (1 pN = one billionth of a Newton) – comparable in magnitude to those that act on proteins and genes in cells. “In principle, we can capture any type of biomolecule with these clamps and investigate its physical properties,” says Tim Liedl.

The effect of the applied force is read out by taking advantage of the phenomenon of Förster Resonant Energy Transfer (FRET). “FRET involves the transfer of energy between two fluorescent dyes and is strongly dependent on the distance between them.” explains Professor Philip Tinnefeld from TU Braunschweig. When the force applied to the test molecule is sufficient to deform it, the distance between the fluorescent markers changes and the magnitude of energy transfer serves as an exquisitely precise measure of the distortion of the test molecule on the nanometer scale.

Together with Dina Grohmann from Universität Regensburg, the team has used the new technique to investigate the properties of the so-called TATA Binding Protein, an important gene regulator which binds to a specific upstream nucleotide sequence in genes and helps to trigger their expression. They found that the TATA protein can no longer perform its normal function if its target sequence is subjected to a force of more than 6 pN. – The new technology has just made its debut. But since the clamps are minuscule and operate autonomously, it may well be possible in the future to use them to study molecular processes in living cells in real time.

Sometimes reading these news releases, my mind is boggled. What an extraordinary time to live.

Here’s a link to and a citation for the paper,

Molecular force spectroscopy with a DNA origami–based nanoscopic force clamp by Philipp C. Nickels, Bettina Wünsch, Phil Holzmeister, Wooli Bae, Luisa M. Kneer, Dina Grohmann, Philip Tinnefeld, Tim Lied. Science  21 Oct 2016: Vol. 354, Issue 6310, pp. 305-307 DOI: 10.1126/science.aah5974

This paper is behind a paywall.

A planet-satellite model for nanoparticles

For anyone who visualizes atoms as planets (many of us were taught to think of atoms and their electrons in that way) then, the planet-satellite model for nanoparticles proposed by scientists at the Nanosystems Initiative Munich (NIM) will have a comforting familiarity, Here’s the model as per a Dec. 13, 2013 news item from Nanowerk,

Nanosystems Initiative Munich (NIM) physicists have developed a “planet-satellite model” to precisely connect and arrange nanoparticles in three-dimensional structures. Like photosystems of plants and algae, the model might in future serve to collect and convert energy.

If the scientists‘ nanoparticles were one million times larger, the laboratory would look like an arts and crafts room at Christmas time: gold, silver and colorful shiny spheres in different sizes and filaments in various lengths. For at the center of the nanoscale “planet-satellite model” there is a gold particle which is orbited by other nanoparticles made of silver, cadmium selenide or organic dyes.

A Dec. 2, 2013 NIM press release, which originated the news item, describes the proposed model in detail,

As if by magic, cleverly designed DNA strands connect the satellites with the central planet in a very precise manner. The technique behind this, called “DNA origami”, is a specialty of physics professor Tim Liedl (LMU Munich) and his team. The expertise on the optical characterization of the individual nanosystems is contributed by Professor Jochen Feldmann, Chair of Photonics and Optoelectronics at LMU and Coordinator of the Nanosystems Initiative Munich (NIM).

Large or small, near or far

A distinctive feature of the new model is the modular assembly system which allows the scientists to modify all aspects of the structure very easily and in a controlled manner: the size of the central nanoparticle, the types and sizes of the “satellites” and the distance between planet and satellite particle. It also enables the physicists to adapt and optimize their system for other purposes.

Artificial photosystem

Metals, semiconductors or fluorescent organic molecules serve as satellites. Thus, like the antenna molecules in natural photosystems, such satellite elements might in future be organized to collect light energy and transfer it to a catalytic reaction center where it is converted into another form of energy. For the time being, however, the model allows the scientists to investigate basic physical effects such as the so-called quenching process, which refers to the changing fluorescence intensity of a dye molecule as a function of the distance to the central gold nanoparticle.

“The modular assembly principle and the high yield we obtained in the production of the planet-satellite systems were the crucial factors for reliably investigating this well-known effect with the new methods,” explains Robert Schreiber, lead author of the study.

A whole new cosmos

In addition, the scientists succeeded in joining individual planet-satellite units together into larger structures, combining them as desired. This way, it might be possible to develop complex and functional three-dimensional nanosystems in future, which could be used as directed energy funnels, in Raman spectroscopy or as nanoporous materials for catalytic applications.

The physicists have supplied an image illustrating their model,

 

[downloaded from http://www.nano-initiative-munich.de/index.php?eID=tx_cms_showpic&file=uploads%2Fpics%2FBasiccover_6_Zeilen_02.jpg&md5=aec790fc11262dc94b41a440fa6788baeacfac97&parameters[0]=YTo0OntzOjU6IndpZHRoIjtzOjQ6IjUwMG0iO3M6NjoiaGVpZ2h0IjtzOjM6IjUw&parameters[1]=MCI7czo3OiJib2R5VGFnIjtzOjI0OiI8Ym9keSBiZ0NvbG9yPSIjZmZmZmZmIj4i&parameters[2]=O3M6NDoid3JhcCI7czozNzoiPGEgaHJlZj0iamF2YXNjcmlwdDpjbG9zZSgpOyI%2B&parameters[3]=IHwgPC9hPiI7fQ%3D%3D] Courtesy NIM

[downloaded from http://www.nano-initiative-munich.de/index.php?eID=tx_cms_showpic&file=uploads%2Fpics%2FBasiccover_6_Zeilen_02.jpg&md5=aec790fc11262dc94b41a440fa6788baeacfac97&parameters[0]=YTo0OntzOjU6IndpZHRoIjtzOjQ6IjUwMG0iO3M6NjoiaGVpZ2h0IjtzOjM6IjUw&parameters[1]=MCI7czo3OiJib2R5VGFnIjtzOjI0OiI8Ym9keSBiZ0NvbG9yPSIjZmZmZmZmIj4i&parameters[2]=O3M6NDoid3JhcCI7czozNzoiPGEgaHJlZj0iamF2YXNjcmlwdDpjbG9zZSgpOyI%2B&parameters[3]=IHwgPC9hPiI7fQ%3D%3D] Courtesy NIM

Here’s a link to and a citation for the researchers’ paper,

Hierarchical assembly of metal nanoparticles, quantum dots and organic dyes using DNA origami scaffolds by Robert Schreiber, Jaekwon Do, Eva-Maria Roller, Tao Zhang, Verena J. Schüller, Philipp C. Nickels, Jochen Feldmann, & Tim Liedl. Nature Nanotechnology (2013) doi:10.1038/nnano.2013.253 Published online 01 December 2013

It is behind a paywall but you can preview it for free via ReadCube Access.