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  1. AU="Kainrath, Stephanie"
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  1. Article ; Online: Design and Application of Light-Regulated Receptor Tyrosine Kinases.

    Kainrath, Stephanie / Janovjak, Harald

    Methods in molecular biology (Clifton, N.J.)

    2020  Volume 2173, Page(s) 233–246

    Abstract: Understanding how the activity of membrane receptors and cellular signaling pathways shapes cell behavior is of fundamental interest in basic and applied research. Reengineering receptors to react to light instead of their cognate ligands allows for ... ...

    Abstract Understanding how the activity of membrane receptors and cellular signaling pathways shapes cell behavior is of fundamental interest in basic and applied research. Reengineering receptors to react to light instead of their cognate ligands allows for generating defined signaling inputs with high spatial and temporal precision and facilitates the dissection of complex signaling networks. Here, we describe fundamental considerations in the design of light-regulated receptor tyrosine kinases (Opto-RTKs) and appropriate control experiments. We also introduce methods for transient receptor expression in HEK293 cells, quantitative assessment of signaling activity in reporter gene assays, semiquantitative assessment of (in)activation time courses through Western blot (WB) analysis, and easy to implement light stimulation hardware.
    MeSH term(s) Blotting, Western ; HEK293 Cells ; Humans ; Optogenetics/methods ; Phosphorylation/genetics ; Phosphorylation/physiology ; Receptor Protein-Tyrosine Kinases/genetics ; Receptor Protein-Tyrosine Kinases/metabolism ; Signal Transduction/genetics ; Signal Transduction/physiology
    Chemical Substances Receptor Protein-Tyrosine Kinases (EC 2.7.10.1)
    Language English
    Publishing date 2020-08-06
    Publishing country United States
    Document type Journal Article
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-0716-0755-8_16
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article: Fähigkeiten und Kompetenzen von Community Health Nurses. Ein Scoping Review

    Lidauer, Harald / Kainrath, Stephanie / Schulc, Eva / Müller, Gerhard / Stummer, Harald

    Pflegewissenschaft

    2022  Volume 24, Issue 4, Page(s) 230

    Language German
    Document type Article
    ZDB-ID 2407183-3
    ISSN 1662-3029
    Database Current Contents Medicine

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 5131: Show issues Location:
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  3. Article: A Light-Oxygen-Voltage Receptor Integrates Light and Temperature

    Dietler, Julia / Schubert, Roman / Krafft, Tobias G.A. / Meiler, Simone / Kainrath, Stephanie / Richter, Florian / Schweimer, Kristian / Weyand, Michael / Janovjak, Harald / Möglich, Andreas

    Journal of molecular biology. 2021 July 23, v. 433, no. 15

    2021  

    Abstract: Sensory photoreceptors enable organisms to adjust their physiology, behavior, and development in response to light, generally with spatiotemporal acuity and reversibility. These traits underlie the use of photoreceptors as genetically encoded actuators ... ...

    Abstract Sensory photoreceptors enable organisms to adjust their physiology, behavior, and development in response to light, generally with spatiotemporal acuity and reversibility. These traits underlie the use of photoreceptors as genetically encoded actuators to alter by light the state and properties of heterologous organisms. Subsumed as optogenetics, pertinent approaches enable regulating diverse cellular processes, not least gene expression. Here, we controlled the widely used Tet repressor by coupling to light-oxygen-voltage (LOV) modules that either homodimerize or dissociate under blue light. Repression could thus be elevated or relieved, and consequently protein expression was modulated by light. Strikingly, the homodimeric RsLOV module from Rhodobacter sphaeroides not only dissociated under light but intrinsically reacted to temperature. The limited light responses of wild-type RsLOV at 37 °C were enhanced in two variants that exhibited closely similar photochemistry and structure. One variant improved the weak homodimerization affinity of 40 µM by two-fold and thus also bestowed light sensitivity on a receptor tyrosine kinase. Certain photoreceptors, exemplified by RsLOV, can evidently moonlight as temperature sensors which immediately bears on their application in optogenetics and biotechnology. Properly accounted for, the temperature sensitivity can be leveraged for the construction of signal-responsive cellular circuits.
    Keywords Rhodobacter sphaeroides ; biotechnology ; blue light ; gene expression ; molecular biology ; optogenetics ; photochemistry ; protein synthesis ; receptor protein-tyrosine kinase ; temperature
    Language English
    Dates of publication 2021-0723
    Publishing place Elsevier Ltd
    Document type Article
    ZDB-ID 80229-3
    ISSN 1089-8638 ; 0022-2836
    ISSN (online) 1089-8638
    ISSN 0022-2836
    DOI 10.1016/j.jmb.2021.167107
    Database NAL-Catalogue (AGRICOLA)

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  4. Article ; Online: Green-Light-Induced Inactivation of Receptor Signaling Using Cobalamin-Binding Domains.

    Kainrath, Stephanie / Stadler, Manuela / Reichhart, Eva / Distel, Martin / Janovjak, Harald

    Angewandte Chemie (International ed. in English)

    2017  Volume 56, Issue 16, Page(s) 4608–4611

    Abstract: Optogenetics and photopharmacology provide spatiotemporally precise control over protein interactions and protein function in cells and animals. Optogenetic methods that are sensitive to green light and can be used to break protein complexes are not ... ...

    Abstract Optogenetics and photopharmacology provide spatiotemporally precise control over protein interactions and protein function in cells and animals. Optogenetic methods that are sensitive to green light and can be used to break protein complexes are not broadly available but would enable multichromatic experiments with previously inaccessible biological targets. Herein, we repurposed cobalamin (vitamin B12) binding domains of bacterial CarH transcription factors for green-light-induced receptor dissociation. In cultured cells, we observed oligomerization-induced cell signaling for the fibroblast growth factor receptor 1 fused to cobalamin-binding domains in the dark that was rapidly eliminated upon illumination. In zebrafish embryos expressing fusion receptors, green light endowed control over aberrant fibroblast growth factor signaling during development. Green-light-induced domain dissociation and light-inactivated receptors will critically expand the optogenetic toolbox for control of biological processes.
    Language English
    Publishing date 2017-03-20
    Publishing country Germany
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2011836-3
    ISSN 1521-3773 ; 1433-7851
    ISSN (online) 1521-3773
    ISSN 1433-7851
    DOI 10.1002/anie.201611998
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: A Light-Oxygen-Voltage Receptor Integrates Light and Temperature.

    Dietler, Julia / Schubert, Roman / Krafft, Tobias G A / Meiler, Simone / Kainrath, Stephanie / Richter, Florian / Schweimer, Kristian / Weyand, Michael / Janovjak, Harald / Möglich, Andreas

    Journal of molecular biology

    2021  Volume 433, Issue 15, Page(s) 167107

    Abstract: Sensory photoreceptors enable organisms to adjust their physiology, behavior, and development in response to light, generally with spatiotemporal acuity and reversibility. These traits underlie the use of photoreceptors as genetically encoded actuators ... ...

    Abstract Sensory photoreceptors enable organisms to adjust their physiology, behavior, and development in response to light, generally with spatiotemporal acuity and reversibility. These traits underlie the use of photoreceptors as genetically encoded actuators to alter by light the state and properties of heterologous organisms. Subsumed as optogenetics, pertinent approaches enable regulating diverse cellular processes, not least gene expression. Here, we controlled the widely used Tet repressor by coupling to light-oxygen-voltage (LOV) modules that either homodimerize or dissociate under blue light. Repression could thus be elevated or relieved, and consequently protein expression was modulated by light. Strikingly, the homodimeric RsLOV module from Rhodobacter sphaeroides not only dissociated under light but intrinsically reacted to temperature. The limited light responses of wild-type RsLOV at 37 °C were enhanced in two variants that exhibited closely similar photochemistry and structure. One variant improved the weak homodimerization affinity of 40 µM by two-fold and thus also bestowed light sensitivity on a receptor tyrosine kinase. Certain photoreceptors, exemplified by RsLOV, can evidently moonlight as temperature sensors which immediately bears on their application in optogenetics and biotechnology. Properly accounted for, the temperature sensitivity can be leveraged for the construction of signal-responsive cellular circuits.
    MeSH term(s) Bacterial Proteins/chemistry ; Bacterial Proteins/metabolism ; Gene Expression Regulation, Bacterial ; Models, Molecular ; Optogenetics ; Oxygen/metabolism ; Protein Domains ; Protein Stability ; Protein Structure, Secondary ; Receptor Protein-Tyrosine Kinases ; Repressor Proteins/genetics ; Repressor Proteins/metabolism ; Rhodobacter sphaeroides/chemistry ; Rhodobacter sphaeroides/metabolism ; Temperature
    Chemical Substances Bacterial Proteins ; Repressor Proteins ; tetracycline resistance-encoding transposon repressor protein ; Receptor Protein-Tyrosine Kinases (EC 2.7.10.1) ; Oxygen (S88TT14065)
    Language English
    Publishing date 2021-06-17
    Publishing country Netherlands
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 80229-3
    ISSN 1089-8638 ; 0022-2836
    ISSN (online) 1089-8638
    ISSN 0022-2836
    DOI 10.1016/j.jmb.2021.167107
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    In stock of ZB MED Cologne/Königswinter

    Zs.A 867: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
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  6. Article ; Online: Optical functionalization of human Class A orphan G-protein-coupled receptors.

    Morri, Maurizio / Sanchez-Romero, Inmaculada / Tichy, Alexandra-Madelaine / Kainrath, Stephanie / Gerrard, Elliot J / Hirschfeld, Priscila P / Schwarz, Jan / Janovjak, Harald

    Nature communications

    2018  Volume 9, Issue 1, Page(s) 1950

    Abstract: G-protein-coupled receptors (GPCRs) form the largest receptor family, relay environmental stimuli to changes in cell behavior and represent prime drug targets. Many GPCRs are classified as orphan receptors because of the limited knowledge on their ... ...

    Abstract G-protein-coupled receptors (GPCRs) form the largest receptor family, relay environmental stimuli to changes in cell behavior and represent prime drug targets. Many GPCRs are classified as orphan receptors because of the limited knowledge on their ligands and coupling to cellular signaling machineries. Here, we engineer a library of 63 chimeric receptors that contain the signaling domains of human orphan and understudied GPCRs functionally linked to the light-sensing domain of rhodopsin. Upon stimulation with visible light, we identify activation of canonical cell signaling pathways, including cAMP-, Ca
    MeSH term(s) Calcium/metabolism ; Cyclic AMP/metabolism ; HEK293 Cells ; Humans ; Light ; Microscopy, Confocal ; Protein Engineering/methods ; Receptors, G-Protein-Coupled/genetics ; Receptors, G-Protein-Coupled/metabolism ; Recombinant Fusion Proteins/genetics ; Recombinant Fusion Proteins/metabolism ; Rhodopsin/genetics ; Rhodopsin/metabolism ; Signal Transduction/genetics ; Signal Transduction/radiation effects
    Chemical Substances Receptors, G-Protein-Coupled ; Recombinant Fusion Proteins ; Rhodopsin (9009-81-8) ; Cyclic AMP (E0399OZS9N) ; Calcium (SY7Q814VUP)
    Language English
    Publishing date 2018-05-16
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2553671-0
    ISSN 2041-1723 ; 2041-1723
    ISSN (online) 2041-1723
    ISSN 2041-1723
    DOI 10.1038/s41467-018-04342-1
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article ; Online: Isolation of synaptic vesicles from genetically engineered cultured neurons.

    McKenzie, Catherine / Spanova, Miroslava / Johnson, Alexander / Kainrath, Stephanie / Zheden, Vanessa / Sitte, Harald H / Janovjak, Harald

    Journal of neuroscience methods

    2018  Volume 312, Page(s) 114–121

    Abstract: Background: Synaptic vesicles (SVs) are an integral part of the neurotransmission machinery, and isolation of SVs from their host neuron is necessary to reveal their most fundamental biochemical and functional properties in in vitro assays. Isolated SVs ...

    Abstract Background: Synaptic vesicles (SVs) are an integral part of the neurotransmission machinery, and isolation of SVs from their host neuron is necessary to reveal their most fundamental biochemical and functional properties in in vitro assays. Isolated SVs from neurons that have been genetically engineered, e.g. to introduce genetically encoded indicators, are not readily available but would permit new insights into SV structure and function. Furthermore, it is unclear if cultured neurons can provide sufficient starting material for SV isolation procedures.
    New method: Here, we demonstrate an efficient ex vivo procedure to obtain functional SVs from cultured rat cortical neurons after genetic engineering with a lentivirus.
    Results: We show that ∼10
    Comparison with existing methods: Obtaining SVs from genetically engineered neurons currently generally requires the availability of transgenic animals, which is constrained by technical (e.g. cost and time) and biological (e.g. developmental defects and lethality) limitations.
    Conclusions: These results demonstrate the modification and isolation of functional SVs using cultured neurons and viral transduction. The ability to readily obtain SVs from genetically engineered neurons will permit linking in situ studies to in vitro experiments in a variety of genetic contexts.
    MeSH term(s) Animals ; Cell Fractionation/methods ; Cells, Cultured ; Genetic Engineering ; Glutamic Acid/metabolism ; Lentivirus/physiology ; Neurons/metabolism ; Rats, Wistar ; Synaptic Vesicles/metabolism
    Chemical Substances Glutamic Acid (3KX376GY7L)
    Language English
    Publishing date 2018-11-26
    Publishing country Netherlands
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 282721-9
    ISSN 1872-678X ; 0165-0270
    ISSN (online) 1872-678X
    ISSN 0165-0270
    DOI 10.1016/j.jneumeth.2018.11.018
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    In stock of ZB MED Cologne/Königswinter

    Zs.A 1486: Show issues Location:
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