Konformationelle Antikörperfragmente für amyloide Erkrankungen(S. 12)
Christian Haupt, Dietmar Rudolf Thal, Uwe Horn, Marcus Fändrich
Conformational diseases, such as Alzheimer’s or Parkinson’s, are characterized by the misfolding of endogenous polypeptide chains into abnormal amyloid fibril conformations. To better analyze this process and its biological consequences and to generate tools for targeted interference in vitro and in vivo, we have biotechnologically generated a set of antibody fragments that are able to discriminate by binding between different amyloid assembly states.
GPCRs als hochrelevante Targets für die Wirkstoffentwicklung(S. 15)
Christine Hiller, Peter Gmeiner
G protein-coupled receptors are involved in numerous physiological processes and provide attractive drug targets for diverse diseases. However, the development of selective, efficacious drugs targeting this family of membrane proteins remains challenging due to multidimensional selectivity profiles of GPCR-agonists and antagonists. Selective GPCR ligands have the potential to induce or stabilize individual receptor conformations or to preferentially bind one particular receptor subtype.
Nanotunnel in bakteriellen Membranen(S. 19)
Porins are pore forming channels in the outer membrane of bacteria, mitochondria, and chloroplasts. Their dominant function is to enable the passive diffusion of nutrient molecules over membranes. Porins can reduce the translocation barrier for larger substrate molecules through specific substrate binding sites as has been demonstrated for specific porins. Unspecific porins are formed by diffusion pores of enlarged dia - meter for the passage of small organic and inorganic molecules and their pore size is often critical for the uptake of antibiotics.
Modulare hybride Wirkstofftransporter auf der Basis bakterieller Toxine(S. 22)
Holger Barth, Tanja Well
The targeted delivery of pharmacologically active molecules into the cytosol of mammalian cells is a central goal of experimental pharmacology and pharmacotherapy. By coupling cell-permeable bacterial protein toxins to streptavidin, we generated modular hybrid protein carriers for specific delivery of biotin-labelled molecules into the cytosol of target cells.
Superinfection exclusion und DNA-Injektion bei Siphoviridae-Phagen(S. 26)
Knut J. Heller, Horst Neve
Superinfection exclusion (SIE) is a mechanism by which a prophage residing in the genome of its host bacterium prevents infection of its host by other phages. During recent years, we have studied superinfection exclusion by prophage TP-J34. Its host is Streptococcus thermophilus J34, a bacterium applied as yoghurt starter. Here we present our data on identifying the SIE protein of TP-J34, its mode of action, X-ray structure, and target protein in the superinfecting phage.
Laser-Mikrodissektion und RNA-Seq zur Analyse der Genexpression in Pilzen(S. 30)
Analysis of gene expression in specialized cells and organs is a powerful method to elucidate mechanisms of cell differentiation. However, in many organisms including fungi, separation of cells of interest from neighbour - ing cells can be difficult. One solution is the use of laser microdissection to cut selected regions from samples under microscopic control. RNA isolated from the harvested cells can be used for RNA-Seq to obtain an overview of global gene expression in the analyzed structures.
Kraftspektroskopie – Dynamik und Manipulation einzelner Moleküle(S. 33)
Gabriel Zoldak, Matthias Rief
Over the last decade, the development of single molecule methods has allowed us to interface individual biomolecules, to subject them to mechanical load and to measure their conformational response. In this article, we introduce the new possibilities of force spectroscopy by AFM (atomic force microscope) and optical tweezers for the study of proteins. Applications of force spectroscopy for protein folding as well as forcedependent ligand binding in cell adhesion are discussed.
Infrarotspektroskopie zur Strukturuntersuchung von Prionen(S. 36)
Martin L. Daus, Michael Beekes, Peter Lasch
The structure of a protein is of major importance for the understanding of its function. Infrared spectroscopy allows the analysis of protein structure and dynamics, even from large proteins or from proteins with poor crystal-forming properties, without the need of sample labelling or staining. This technique is relatively simple in use and can provide data within a very short time. Therefore, infrared spectroscopy belongs to the methods of choice to study the structure of prions.
Modellierung metabolischer Netzwerke im menschlichen Körper(S. 39)
Henrik Cordes, Christoph Thiel, Lars M. Blank, Lars Küpfer
Human physiology covers multiple levels of biological organization. For a mechanistic understanding of diseases and a rational development of targeted therapeutic strategies, a simultaneous consideration of processes at the cellular scale and at the whole-body level is desirable. We here discuss how metabolic network models can be integrated into physiologybased pharmacokinetic models at organism level. Several potential applications involving both endogenous and exogenous metabolism are given.
Schnelle Detektion mikrobieller Kontaminationen in Lebensmitteln(S. 43)
Jan Lenke, Helmut Erdmann
An important issue in food industries is the detection of microbial contaminants. Flow cytometry not only allows the rapid detection of individual cells but at the same time the quantification and determination of their viability. The concept of flow cytometric analysis has been realized by a miniaturized integrated lab-on-a-chip setup. A microfluidic system using hydrodynamic focusing was developed on a chip.
Entzündung ohne Erreger – Rolle des Immunsystems nach Schlaganfall(S. 48)
Daniel-Christoph Wagner, Karoline Möller, Claudia Pösel
Cerebral stroke is one of the most important causes for death and dis - ability. Despite its considerable socio-economic burden, causal therapy is available for only 20 percent of patients. It has been shown that poststroke immune responses significantly influence neurological outcome. Understanding these mechanisms could unveil novel targets for stroke treatment. Here we describe the use of flow cytometry for studying poststroke sterile brain inflammation.
Selbstheilung und potenzielle Unsterblichkeit bei Planarien(S. 51)
Henning Schmitz, Martin Stehling, Luca Gentile
Flow cytometry is by far the most sophisticated and accurate method for sorting of living stem cells. In general, the target cells need to be labeled for various cell identity markers, which are exposed on the surface of the cells. However, in non-model organisms, we usually lack specific labels for such cell surface markers. Here we describe a method for isolating stem cells from planarians with flow cytometry, based on physiological and morphological properties of these cells.
Anwendungen & Produkte
Sicherung der Produktqualität rekombinanter Wachstumsfaktoren(S. 64)
Caroline Blumer Toti, Volker Nölle
The demand for high grade recombinant proteins is growing. Especially in the field of cell-based therapies, all materials used, including cytokines and growth factors must comply to the highly demanding GMP specifications. We show how essential protein parameters are monitored to ensure consistent high product quality using the cytokine interleukin-4 as an example.
Schnelle und reproduzierbare Proteinanalyse(S. 67)
Proteins are analysed in many research and development areas, e. g. the search for biomarkers, and the pharmacological development. The standard method is tryptic digestion which disassembles the proteins in their peptides before they are determined by mass spectrometry. The biggest influence on the reproducibility and speed of the analysis has the sample preparation. Therefore, a new online trypsin digestion platform automates the sample preparation, reduces the analysis duration from hours to minutes, and achieves high reproducibility.
Zellfreie Proteinexpression für Forschung und Produktion(S. 70)
Caroline End, Christian Walczuch, Matthias Buntru
Cell-free protein synthesis (CFPS) is an important tool for molecular bio - logy and can be used in applied and basic research. Especially as a high throughput technology for functional genomics and proteomics, CFPS systems offer a significant advantage compared to living cells. Toxic, membrane or viral proteins can be readily expressed in vitro. However, a low synthesis performance currently limits the use for bioproduction purposes. To overcome this limitation, new systems are being developed.
Biokatalyse in unkonventionellen Reaktionsmedien(S. 100)
Frank Hollmann, Selin Kara
Biocatalysis in organic media enables higher concentrations of mostly hydrophobic reagents compared to traditional aqueous reaction systems. The resulting economic and ecologic advantages call for more intensive research efforts to attain deeper understanding of the behaviour of biocatalysts under non-conventional reaction conditions.
Geobiotechnische Metallgewinnung(S. 103)
Sabrina Hedrich, Axel Schippers
Biomining is the extraction of metals from ores by microorganisms which to date is industrially applied to recover metals (i. e. Cu, Au, Co, Ni) from sulfide ores. Recent research & development has brought about a novel method for the dissolution of oxide ores and promises potential in bio - leaching of waste material (dump material, electronic waste, etc.). Similar microbial processes have been developed for the selective recovery of metals from acidic mine and process waters.