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Ex vivo processing for maturation of Arabidopsis KDEL-tailed cysteine endopeptidase 2 (AtCEP2) pro-enzyme and its storage in endoplasmic reticulum derived organelles

Ex vivo processing for maturation of Arabidopsis KDEL-tailed cysteine endopeptidase 2 (AtCEP2) pro-enzyme and its storage in endoplasmic reticulum derived organelles

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HIERL, Georg, Timo HÖWING, Erika ISONO, Friedrich LOTTSPEICH, Christine GIETL, 2014. Ex vivo processing for maturation of Arabidopsis KDEL-tailed cysteine endopeptidase 2 (AtCEP2) pro-enzyme and its storage in endoplasmic reticulum derived organelles. In: Plant Molecular Biology. 84(6), pp. 605-620. ISSN 0167-4412. eISSN 1573-5028

@article{Hierl2014-04proce-38201, title={Ex vivo processing for maturation of Arabidopsis KDEL-tailed cysteine endopeptidase 2 (AtCEP2) pro-enzyme and its storage in endoplasmic reticulum derived organelles}, year={2014}, doi={10.1007/s11103-013-0157-6}, number={6}, volume={84}, issn={0167-4412}, journal={Plant Molecular Biology}, pages={605--620}, author={Hierl, Georg and Höwing, Timo and Isono, Erika and Lottspeich, Friedrich and Gietl, Christine} }

<rdf:RDF xmlns:rdf="http://www.w3.org/1999/02/22-rdf-syntax-ns#" xmlns:bibo="http://purl.org/ontology/bibo/" xmlns:dc="http://purl.org/dc/elements/1.1/" xmlns:dcterms="http://purl.org/dc/terms/" xmlns:xsd="http://www.w3.org/2001/XMLSchema#" > <rdf:Description rdf:about="https://kops.uni-konstanz.de/rdf/resource/123456789/38201"> <dc:contributor>Gietl, Christine</dc:contributor> <dc:creator>Lottspeich, Friedrich</dc:creator> <dc:language>eng</dc:language> <dc:contributor>Höwing, Timo</dc:contributor> <dc:contributor>Lottspeich, Friedrich</dc:contributor> <dc:creator>Hierl, Georg</dc:creator> <dc:contributor>Hierl, Georg</dc:contributor> <bibo:uri rdf:resource="https://kops.uni-konstanz.de/handle/123456789/38201"/> <dcterms:issued>2014-04</dcterms:issued> <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2017-03-29T13:48:27Z</dc:date> <dc:contributor>Isono, Erika</dc:contributor> <dcterms:rights rdf:resource="http://nbn-resolving.de/urn:nbn:de:bsz:352-20150914100631302-4485392-8"/> <dcterms:abstract xml:lang="eng">Ricinosomes are specialized ER-derived organelles that store the inactive pro-forms of KDEL-tailed cysteine endopeptidases (KDEL-CysEP) associated with programmed cell death (PCD). The Arabidopsis genome encodes three KDEL-CysEP (AtCEP1, AtCEP2, and AtCEP3) that are differentially expressed in vegetative and generative tissues undergoing PCD. These Arabidopsis proteases have not been characterized at a biochemical level, nor have they been localized intracellularly. In this study, we characterized AtCEP2. A 3xHA-mCherry-AtCEP2 gene fusion including pro-peptide and KDEL targeting sequences expressed under control of the endogenous promoter enabled us to isolate AtCEP2 "ex vivo". The purified protein was shown to be activated in a pH-dependent manner. After activation, however, protease activity was pH-independent. Analysis of substrate specificity showed that AtCEP2 accepts proline near the cleavage site, which is a rare feature specific for KDEL-CysEPs. mCherry-AtCEP2 was detected in the epidermal layers of leaves, hypocotyls and roots; in the root, it was predominantly found in the elongation zone and root cap. Co-localization with an ER membrane marker showed that mCherry-AtCEP2 was stored in two different types of ER-derived organelles: 10 μm long spindle shaped organelles as well as round vesicles with a diameter of approximately 1 μm. The long organelles appear to be ER bodies, which are found specifically in Brassicacae. The round vesicles strongly resemble the ricinosomes first described in castor bean. This study provides a first evidence for the existence of ricinosomes in Arabidopsis, and may open up new avenues of research in the field of PCD and developmental tissue remodeling.</dcterms:abstract> <dc:creator>Höwing, Timo</dc:creator> <dcterms:title>Ex vivo processing for maturation of Arabidopsis KDEL-tailed cysteine endopeptidase 2 (AtCEP2) pro-enzyme and its storage in endoplasmic reticulum derived organelles</dcterms:title> <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2017-03-29T13:48:27Z</dcterms:available> <dc:creator>Gietl, Christine</dc:creator> <dc:creator>Isono, Erika</dc:creator> </rdf:Description> </rdf:RDF>

Dateiabrufe seit 29.03.2017 (Informationen über die Zugriffsstatistik)

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