Interference Probe of Plant Endoplasmic Reticulum

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The plant endomembrane system consists of several distinct and specialized compartments, including the endoplasmic reticulum (ER), the Golgi, the vacuole, and endosomal compartments. Together, they form a highly dynamic, fundamental, and interconnected system responsible for the synthesis, transport, degradation, and segregation of macromolecules. In multicellular organisms, traditional genetic approaches are challenging to anatomize endomembrane function due to the redundancy and importance of genes, making complementary small molecule approaches attractive.

Lifeasible is a leading provider of comprehensive, high-quality services for plant sciences. We provide an interference probe of plant endoplasmic reticulum for research purposes to customers worldwide. We look forward to working with our customers to discover the secrets of life sciences.

Establishment of Interference Probe of Plant Endoplasmic Reticulum

Step	Methods
Development of Small Molecule Disrupter for Endoplasmic Reticulum Structure	To perturb the ER / Golgi structure, Lifeasible performs microscopic screening of small molecule inhibitors for the plant ER by using a GFP marker. At the same time, we analyze commercially available analogs of the initial hit molecule to discover an analog with improved potency that can inhibit plant growth. In addition, the significant effect of this small molecule on ER structure transforms the ER from a typical tubular pool to a predominantly vesicular structure composed of extensive aggregations of vesicles of different sizes and shapes.
In vivo Protein Labeling with Clickable Photoaffinity Labeling Prob	To identify cellular targets for small molecule disrupters, we design both bioactive and "dead" clickable photoaffinity probes that we can use to identify proteins selectively labeled by the bioactive probes following Cu⁺ -catalyzed azide-alkye cycloaddition. We synthesize clickable photoaffinity probes by attaching the required aryl azide and alkyne functional groups at appropriate locations of the small molecule disrupters.
Clickable Photoaffinity Probe Labels Oleosins	Oleosins are proteins that stabilize lipid droplets (LDs) in plants, which are required to establish early ER structure in seeds and seedlings. However, if oleosins change in the plant cells, the structure of the ER will also change.

Function Discovery of Endoplasmic Reticulum with Interference Probe

Fig.1. ER function is explored with a clickable ER interference probe. (Dejonghe W., et al., 2021)

In conclusion, we identify a novel molecule that perturbs ER structure and further synthesizes the clickable photoaffinity labeling probes. This probe selectively labels the oleaginous proteins in plants to realize the labeling of ER structure and the exploration of ER function.
We explore the effect of local ER on plants by using the interference probe in local areas of plants.
Likewise, we also use the interference probe at a specific time of plant growth to explore the influence of ER on plants at a certain stage.

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Reference

Dejonghe W., et al. (2021). "In Planta Labeling Using a Clickable ER-Disrupting Probe Suggests a Role for Oleosins in Arabidopsis Seedling ER Integrity." ACS Chem Biol. 16 (11), 2151-2157.

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