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Auxin profiling in Arabidopsis endoplasmic reticulum.

ABSTRACT: Endoplasmic reticulum (ER) is an intracellular extensive network of membranes. Among major ER functions belong proteosynthesis, protein folding, their post-transcriptional modification and sorting within the cell and last but not least lipid anabolism. Moreover, several recent studies have indicated involvement of plant ER in regulation of intracellular auxin homeostasis by modulation of its metabolism. Therefore, in order to study auxin metabolome in ER, it is necessary to obtain highly enriched and, if possible, pure ER fraction. The separation of ER is troublesome due to similar biochemical properties with other cellular endomembranes. The majority of published protocols for ER isolation utilise density gradient ultracentrifugation despite the suboptimal resolving power of this approach. This work aims to optimize the isolation of ER from Arabidopsis thaliana for the subsequent determination of ER-specific auxin metabolite profiles by mass spectrometry. Following auxin metabolite determination showed a high increase of active auxin form within ER compared to whole plant. Moreover, our optimized isolation of ER may be further followed by multiple “omics” technologies including analysis of macromolecular as well as low molecular-weight compound from same sample.

INSTRUMENT(S): maXis

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Plant Cell, Seedling

SUBMITTER: René Lenobel

LAB HEAD: Aleš Pěnčík

PROVIDER: PXD027522 | Pride | 2021-11-09

REPOSITORIES: Pride

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