Ruben Garrecht

Karlsruhe Institute of Technology

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Christof M. Niemeyer

Karlsruhe Institute of Technology

Michael Hirtz

Karlsruhe Institute of Technology

Simone Weigel

Karlsruhe Institute of Technology

Ravi Kumar

Karlsruhe Institute of Technology

Dominic Kamps

Max Planck Institute of Molecular Physiology

Leif Dehmelt

TU Dortmund University

Muthukumaran Venkatachalapathy

Ben-Gurion University of the Negev

Michael M. Orlich

University Medical Center Hamburg-Eppendorf

Yao‐Wen Wu

Umeå University

Rebecca Meyer

California Baptist University

Cooperative Institutions

TU Dortmund University

Karlsruhe Institute of Technology

Max Planck Institute of Molecular Physiology

Leibniz Research Centre for Working Environment and Human Factors

University of Duisburg-Essen

Nanyang Technological University

University of Freiburg

King's College London

City University of Hong Kong

Max Planck Society

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Solid‐Phase Synthesis and Purification of Protein–DNA Origami Nanostructures

Chemistry - A European Journal (2019)

Teresa Burgahn Ruben Garrecht Kersten S. Rabe Christof M. Niemeyer

Abstract Invited for the cover of this issue are C. M. Niemeyer and co‐workers at the Karlsruhe Institute of Technology (KIT). The cover picture shows a comic hero with magnetic superpowers who pulls pure, protein‐decorated DNA origami nanostructures out of a crude reaction mixture. Read the full text of the article at 10.1002/chem.201805506 .

HERO

10.1002/chem.201900572

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Front Cover: Solid‐Phase Synthesis and Purification of Protein–DNA Origami Nanostructures (Chem. Eur. J. 14/2019)

Chemistry - A European Journal (2019)

Teresa Burgahn Ruben Garrecht Kersten S. Rabe Christof M. Niemeyer

Similar to a comic hero with magnetic superpowers, a microbead assay allows for the efficient extraction of protein-decorated DNA origami nanostructures from crude reaction mixtures. The method provides high yields and delivers pure samples to enable direct AFM analysis for the evaluation of orthogonal immobilization strategies. More information can be found in the Communication by C. M. Niemeyer et al. on page 3483.

Microbead (research)

DNA origami

Front cover

HERO

10.1002/chem.201900571

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“Molecular Activity Painting”: schaltbare, lichtgesteuerte Manipulation in lebenden Zellen

Angewandte Chemie (2017)

Xi Chen Muthukumaran Venkatachalapathy Dominic Kamps Simone Weigel Ravi Kumar

Die akute subzelluläre Rekrutierung von Proteinen ist eine leistungsfähige Methode zur Untersuchung biologischer Netzwerke. Hoch-dynamische Signalprozesse an der Plasmamembran können bisher jedoch nur bedingt räumlich und zeitlich untersucht werden. Insbesondere durch laterale Diffusion an der Plasmamembran ist eine anhaltende lokale Steuerung molekularer Funktionen schwer zu realisieren. Hier wird die “Molekulare-Aktivitäts-Malerei” (Molecular Activity Painting, MAP) vorgestellt, eine neuartige Technik, die photoaktivierbare chemisch induzierte Dimerisierung (pCID) mit immobilisierten künstlichen Rezeptoren kombiniert. Die Immobilisierung der künstlichen Rezeptoren durch oberflächengebundene Antikörper hemmt die laterale Diffusion. Hierdurch wird schnelles und stabiles Malen (Painting) von Signalmolekülen und deren Aktivität an der Plasmamembran im Mikrometermaßstab ermöglicht. Stabiles Malen des RhoA-Myosin-Aktivators GEF-H1 kann kontraktile Actomyosin-Strukturen in lebenden Zellen induzieren.

10.1002/ange.201611432

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Citations (14)

Multiscale Origami Structures as Interface for Cells

Angewandte Chemie (2015)

Alessandro Angelin Simone Weigel Ruben Garrecht Rebecca Meyer J. Bauer

Abstract A DNA‐based platform was developed to address fundamental aspects of early stages of cell signaling in living cells. By site‐directed sorting of differently encoded, protein‐decorated DNA origami structures on DNA microarrays, we combine the advantages of the bottom‐up self‐assembly of protein–DNA nanostructures and top‐down micropatterning of solid surfaces to create multiscale origami structures as interface for cells (MOSAIC). In a proof‐of‐principle, we use this technology to analyze the activation of epidermal growth factor (EGF) receptors in living MCF7 cells using DNA origami structures decorated on their surface with distinctive nanoscale arrangements of EGF ligand entities. MOSAIC holds the potential to present to adhered cells well‐defined arrangements of ligands with full control over their number, stoichiometry, and precise nanoscale orientation. It therefore promises novel applications in the life sciences, which cannot be tackled by conventional technologies.

DNA origami

Micropatterning

Interface (matter)

DNA nanotechnology

10.1002/ange.201509772

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Citations (16)

Solid‐Phase Synthesis and Purification of Protein–DNA Origami Nanostructures

Chemistry - A European Journal (2019)

Teresa Burgahn Ruben Garrecht Kersten S. Rabe Christof M. Niemeyer

Abstract We present a facile method for the combined synthesis and purification of protein‐decorated DNA origami nanostructures (DONs). DONs bearing reductively cleavable biotin groups in addition to ligands for ligation of recombinant proteins are bound to magnetic beads. Protein immobilization is conducted with a large protein excess to achieve high ligation yields. Subsequent to cleavage from the solid support, pure sample solutions are obtained which are suitable for direct AFM analysis of occupation patterns. We demonstrate the method's utility using three different orthogonal ligation methods, the “halo‐based oligonucleotide binder” (HOB), a variant of Halo‐tag, the “SpyTag/SpyCatcher” (ST/SC) system, and the enzymatic “ybbR tag” coupling. We find surprisingly low efficiency for ST/SC ligation, presumably due to electrostatic repulsion and steric hindrance, whereas the ybbR method, despite its ternary nature, shows good ligation yields. Our method is particularly useful for the development of novel ligation methods and the synthesis of mechanically fragile DONs that present protein patterns for surface‐based cell assays.

Solid-Phase Synthesis

Cleavage (geology)

10.1002/chem.201805506

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Citations (17)

Inside Back Cover: “Molecular Activity Painting”: Switch-like, Light-Controlled Perturbations inside Living Cells (Angew. Chem. Int. Ed. 21/2017)

Angewandte Chemie International Edition (2017)

Xi Chen Muthukumaran Venkatachalapathy Dominic Kamps Simone Weigel Ravi Kumar

People all around the world carve their initials into trees as a symbol of eternal love. But those carvings are about 1 million times larger than the paintings that were made in living cells by Y.-W. Wu, L. Dehmelt, and co-workers in their Communication on page 5916 ff. Although these cell paintings were not made for eternity, they lasted for several hours––a very long time in the dynamic environment of a living cell. The paintings were made not just for fun, but were used to manipulate cell function to reach a better understanding of complex disease processes such as cancer metastasis.

Eternity

Symbol (formal)

Living cell

Bit (key)

Cell function

10.1002/anie.201703450

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Designed DNA Surfaces for in Vitro Modulation of Natural Killer Cells

ChemBioChem (2016)

Ruben Garrecht Rebecca Meyer Janine Duppach Simone Reipschläger Carsten Watzl

Abstract Natural killer (NK) cells are at the junction of the innate and the adaptive immune response and play a very important role in host defense against viral infections and cancer. They have numerous cell surface receptors that activate or inhibit various intracellular signaling cascades that are then integrated to determine the functional activity of these cells. Here we present a surface‐based approach that aims to tackle the largely unknown molecular mechanisms of signal integration. We use DNA microarrays containing capture oligonucleotides for the DNA‐directed immobilization (DDI) of oligonucleotide‐tagged αCD16 antibodies as ligands for NK cells. We demonstrate that the resulting surfaces can be gradually tuned in terms of ligand density to trigger the activation of living NK cells, as evidenced by degranulation, the release of cytokines, and intracellular Ca 2+ flux, measured at the level of single cells.

Aptamer

CD16

10.1002/cbic.201500629

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Citations (11)

“Molecular Activity Painting”: Switch‐like, Light‐Controlled Perturbations inside Living Cells

Angewandte Chemie International Edition (2017)

Xi Chen Muthukumaran Venkatachalapathy Dominic Kamps Simone Weigel Ravi Kumar

Abstract Acute subcellular protein targeting is a powerful tool to study biological networks. However, signaling at the plasma membrane is highly dynamic, making it difficult to study in space and time. In particular, sustained local control of molecular function is challenging owing to the lateral diffusion of plasma membrane targeted molecules. Herein we present “molecular activity painting” (MAP), a novel technology which combines photoactivatable chemically induced dimerization (pCID) with immobilized artificial receptors. The immobilization of artificial receptors by surface‐immobilized antibodies blocks lateral diffusion, enabling rapid and stable “painting” of signaling molecules and their activity at the plasma membrane with micrometer precision. Using this method, we show that painting of the RhoA‐myosin activator GEF‐H1 induces patterned acto‐myosin contraction inside living cells.

Cell surface receptor

10.1002/anie.201611432

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Citations (41)

Innenrücktitelbild: “Molecular Activity Painting”: schaltbare, lichtgesteuerte Manipulation in lebenden Zellen (Angew. Chem. 21/2017)

Angewandte Chemie (2017)

Xi Chen Muthukumaran Venkatachalapathy Dominic Kamps Simone Weigel Ravi Kumar

Menschen auf der ganzen Welt schnitzen ihre Initialen in Bäume als Zeichen ewiger Liebe. Diese Schnitzereien sind eine Million Mal größer als die Malereien in lebenden Zellen, die Y.-W. Wu, L. Dehmelt und Mitarbeiter in ihrer Zuschrift auf S. 6010 ff. vorstellen. Und auch wenn diese Zellmalereien nicht für die Ewigkeit sind, halten sie doch mehrere Stunden – eine sehr lange Zeit in der dynamischen Umgebung einer lebenden Zelle. Die Malereien dienen nicht dem bloßen Vergnügen, sondern werden zur Manipulation von Zellfunktionen genutzt, um damit ein besseres Verständnis von komplexen Krankheitsprozessen wie Krebsmetastase zu gewinnen.

10.1002/ange.201703450

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Citations (1)

Multiscale Origami Structures as Interface for Cells

Angewandte Chemie International Edition (2015)

Alessandro Angelin Simone Weigel Ruben Garrecht Rebecca Meyer J. Bauer

DNA origami

Micropatterning

Interface (matter)

10.1002/anie.201509772

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Citations (92)