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Publikationen - Molekulare Signalverarbeitung

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Hellmuth, A.; Calderón Villalobos, L. I. A. Radioligand Binding Assays for Determining Dissociation Constants of Phytohormone Receptors (Lois, L. M.; Matthiesen, R. ). Meth. Mol. Biol 1450, 23-34, (2016) ISBN: 978-1-4939-3757-8 DOI: 10.1007/978-1-4939-3759-2_3

In receptor–ligand interactions, dissociation constants provide a key parameter for characterizing binding. Here, we describe filter-based radioligand binding assays at equilibrium, either varying ligand concentrations up to receptor saturation or outcompeting ligand from its receptor with increasing concentrations of ligand analogue. Using the auxin coreceptor system, we illustrate how to use a saturation binding assay to determine the apparent dissociation constant (K D ′ ) for the formation of a ternary TIR1–auxin–AUX/IAA complex. Also, we show how to determine the inhibitory constant (K i) for auxin binding by the coreceptor complex via a competition binding assay. These assays can be applied broadly to characterize a one-site binding reaction of a hormone to its receptor.
Publikation

Guranowski, A.; Miersch, O.; Staswick, P.E.; Suza, W.; Wasternack, C. Substrate specificity and products of side-reactions catalyzed by jasmonate:amino acid synthetase (JAR1) FEBS Letters 581, 815-820, (2007)

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Publikation

Schilling, S.; Stenzel, I.; von Bohlen, A.; Wermann, M.; Schulz, K.; Demuth, H.-U.; Wasternack, C. Isolation and characterization of the glutaminyl cyclases from <i>Solanum tuberosum</i> and <i>Arabidopsis thaliana</i>: implications for physiological functions Biol. Chem 388, 145-153, (2007)

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Publikation

Wasternack, C.; Stenzel, I.; Hause, B.; Hause, G.; Kutter, C.; Maucher, H.; Neumerkel, J.; Feussner, I.; Miersch, O. The wound response in tomato - Role of jasmonic acid J. Plant Physiol 163, 297-306 , (2006)

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Publikation

Flores, R.; Delgado, S.; Gas, M.E.; Carbonell, A.; Molina, D.; Gago, S.; de la Peña, M. Viroids: the minimal non-coding RNA's with autonomous replication FEBS Letters 567, 42-48, (2004)

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Publikation

Morgan, K.E.; Zarembinski, T.I.; Theologis, A.; Abel, S. Biochemical characterization of recombinant polypeptides corresponding to the predicted ßαα-fold in Aux/IAA proteins FEBS Letters 454, 283-287, (1999)

The plant hormone indoleacetic acid (IAA or auxin) transcriptionally activates a select set of early genes. The Auxl IAA class of early auxin-responsive genes encodes a large family of short-lived, nuclear proteins. Aux/IAA polypeptides homo-and heterodimerize, and interact with auxin-response transcription factors (ARFs) via C-terminal regions conserved in both protein families. This shared region contains a predicted βαα motif similar to the prokaryotic β-Ribbon DNA binding domain, which mediates both protein dimerization and DNA recognition. Here, we show by circular dichroism spectroscopy and by chemical cross-linking experiments that recombinant peptides corresponding to the predicted βαα region of three Aux/IAA proteins from Arabidopsis thaliana contain substantial α-helical secondary structure and undergo homo- and heterotypic interactions in vitro. Our results indicate a similar biochemical function of the plant βαα domain and suggest that the βαα fold plays an important role in mediating combinatorial interactions of Aux/IAA and ARF proteins to specifically regulate secondary gene expression in response to auxin.
Publikation

Bohlmann, H.; Vignutelli, A.; Hilpert, B.; Miersch, O.; Wasternack, C.; Apel, K. Wounding and chemicals induce expression of the Arabidopsis gene Thi2.1, encoding a fungal defense thionin, via the octadecanoid pathway FEBS Letters 437, 281-286, (1998)

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Publikation

Hertel, S.; Knöfel, H.-D.; Kramell, R.; Miersch, O. Partial purification and characterization of a jasmonic acid conjugate cleaving amidohydrolase from the fungus <EM>Botryodiplodia theobromae</EM> FEBS Letters 407, 105-110, (1997)

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Publikation

Görschen, E.; Dunaeva, M.; Reeh, I.; Wasternack, C. Overexpression of the jasmonate inducible 23 kDa protein (JIP 23) from barley in transgenic tobacco leads to the repression of leaf proteins FEBS Letters 419, 58-62, (1997)

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Publikation

Feussner, I.; Kühn, H.; Wasternack, C. Hypothesis. Do specific linoleate 13-lipoxygenases initiate b-oxidation? FEBS Letters 406, 1-5, (1997)

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