Endoplasmic reticulum

Dashek, William V.

doi:10.1002/9781118924846.ch3

Cited by 1 publication

(1 citation statement)

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“…Additionally, in plant cells the ER mediates communication between the endomembrane system and non-secretory organelles, such as mitochondria, peroxisomes, and chloroplasts. Proteins in the ER undergo post-translational modifications, such as Nand O-glycosylation and hydrogen bond formation [1][2][3][4]. Studies conducted during the last two decades have also shown that the ER is involved in regulating the distribution of phytohormones and signalling via auxins [5][6][7][8][9], cytokinins [10,11], and ethylene [12].…”

Section: Introductionmentioning

confidence: 99%

Auxin Metabolome Profiling in the Arabidopsis Endoplasmic Reticulum Using an Optimised Organelle Isolation Protocol

Včelařová

Skalický

Chamrád

et al. 2021

IJMS

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The endoplasmic reticulum (ER) is an extensive network of intracellular membranes. Its major functions include proteosynthesis, protein folding, post-transcriptional modification and sorting of proteins within the cell, and lipid anabolism. Moreover, several studies have suggested that it may be involved in regulating intracellular auxin homeostasis in plants by modulating its metabolism. Therefore, to study auxin metabolome in the ER, it is necessary to obtain a highly enriched (ideally, pure) ER fraction. Isolation of the ER is challenging because its biochemical properties are very similar to those of other cellular endomembranes. Most published protocols for ER isolation use density gradient ultracentrifugation, despite its suboptimal resolving power. Here we present an optimised protocol for ER isolation from Arabidopsis thaliana seedlings for the subsequent mass spectrometric determination of ER-specific auxin metabolite profiles. Auxin metabolite analysis revealed highly elevated levels of active auxin form (IAA) within the ER compared to whole plants. Moreover, samples prepared using our optimised isolation ER protocol are amenable to analysis using various “omics” technologies including analyses of both macromolecular and low molecular weight compounds from the same sample.

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