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Publikationen - Molekulare Signalverarbeitung

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Publikation

Ziegler, J.; Abel S. Analysis of amino acids by HPLC/electrospray negative ion tandem mass spectrometry using 9-fluorenylmethoxycarbonyl chloride (Fmoc-Cl) derivatization Amino Acids 46, 2799-2808, (2014) DOI: 10.1007/s00726-014-1837-5

A new method for the determination of amino acids is presented. It combines established methods for the derivatization of primary and secondary amino groups with 9-fluorenylmethoxycarbonyl chloride (Fmoc-Cl) with the subsequent amino acid specific detection of the derivatives by LC–ESI–MS/MS using multiple reaction monitoring (MRM). The derivatization proceeds within 5 min, and the resulting amino acid derivatives can be rapidly purified from matrix by solid-phase extraction (SPE) on HR-X resin and separated by reversed-phase HPLC. The Fmoc derivatives yield several amino acid specific fragment ions which opened the possibility to select amino acid specific MRM transitions. The method was applied to all 20 proteinogenic amino acids, and the quantification was performedusing l-norvaline as standard. A limit of detection as low as 1 fmol/μl with a linear range of up to 125 pmol/μl could be obtained. Intraday and interday precisions were lower than10 % relative standard deviations for most of the amino acids. Quantification usingl-norvaline as internal standard gave very similar results compared to the quantificationusing deuterated amino acid as internal standards. Using this protocol, it was possible to record the amino acid profiles of only a single root from Arabidopsis thaliana seedlings and to compare it with the amino acid profiles of 20 dissected root meristems (200 μm).
Publikation

Wasternack, C.; Hause, B. Jasmonates: biosynthesis, perception, signal transduction and action in plant stress response, growth and development. An update to the 2007 review in <span>Annals of Botany</span> Annals of Botany 111, 1021-1058, (2013) DOI: 10.1093/aob/mct067

Background: Jasmonates are important regulators in plant responses to biotic and abiotic stresses as well as indevelopment. Synthesized from lipid-constituents, the initially formed jasmonic acid is converted to differentmetabolites including the conjugate with isoleucine. Important new components of jasmonate signalling includingits receptor were identified, providing deeper insight into the role of jasmonate signalling pathways in stressresponses and development.Scope: The present review is an update of the review on jasmonates published in this journal in 2007. New dataof the last five years are described with emphasis on metabolites of jasmonates, on jasmonate perception andsignalling, on cross-talk to other plant hormones and on jasmonate signalling in response to herbivores and pathogens,in symbiotic interactions, in flower development, in root growth and in light perception.Conclusions: The last few years have seen breakthroughs in the identification of JASMONATE ZIM DOMAIN(JAZ) proteins and their interactors such as transcription factors and co-repressors, and the crystallization of thejasmonate receptor as well as of the enzyme conjugating jasmonate to amino acids. Now, the complex nature ofnetworks of jasmonate signalling in stress responses and development including hormone cross-talk can beaddressed.
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