What does Tunicamycin do to cells?
Tunicamycin (Tun), a naturally occurring antibiotic, induces ER stress in cells by inhibiting the first step in the biosynthesis of N-linked glycans in the proteins resulting many misfolded proteins .
How does thapsigargin induce ER stress?
ER stress can be induced by treating cells with 0.1–1 µM of thapsigargin for 5 hours. Brefeldin A inhibits transport of proteins from the ER to the Golgi and induces retrograde protein transport from the Golgi apparatus to the endoplasmic reticulum. This leads to the accumulation of unfolded proteins in the ER.
How do you dissolve Tunicamycin?
Tunicamycin is sparingly soluble in aqueous buffers. For maximum solubility in aqueous buffers, tunicamycin should first be dissolved in DMSO and then diluted with the aqueous buffer of choice. Tunicamycin has a solubility of approximately 0.25 mg/ml in a 1:3 solution of DMSO:PBS (pH 7.2) using this method.
What is another name for thapsigargin?
|Preferred IUPAC name (3S,3aR,4S,6S,6aR,7S,8S,9bS)-3,3a-Dihydroxy-3,6,9-trimethyl-2-oxo-2,3,3a,4,5,6,6a,7,8,9b-decahydroazuleno[4,5-b]furan-4,6,7,8-tetrayl 6-acetate 4-butanoate 8-[(2Z)-2-methylbut-2-enoate] 7-octanoate|
|3D model (JSmol)||Interactive image|
How do you detect ER stress?
Generally ER stress is detected indirectly by measuring the levels of specific UPR factors. Commonly measured indicators of UPR activation include; phosphorylated PERK, phosphorylated eIF2α, Gadd153/CHOP, ATF4, Grp78/BiP, Grp94, calreticulin, and protein disulphide isomerase (PDI).
What’s the chemical used to induce ER stress?
Three chemicals are generally used to experimentally induce ER stress: tunicamycin (Sigma), thapsigargin (Sigma), and Brefeldin A (BFA) (Sigma). Although these chemicals target different components of the ER, their common effect is to interfere with ER functions and thereby lead to ER protein misfolding.
What are the two types of glycosylation?
The Notch proteins go through these organelles in their maturation process and can be subject to different types of glycosylation: N-linked glycosylation and O-linked glycosylation (more specifically: O-linked glucose and O-linked fucose).
What is the effect of Tunicamycin on GC cells?
As a potent inhibitor of glycosylation, tunicamycin (Tu) has shown marked antitumor activities in various cancers. In the present study, we attempted to determine the exact effect of Tu on the chemoresistance of GC. The cytotoxic effects of drugs on GC cells were evaluated by cell viability assays, and apoptosis was detected by flow cytometry.
How does tunicamycin work in the endoplasmic reticulum?
Initially identified as a natural antibiotic, tunicamycin (Tu) is also a canonical compound for blocking N-linked glycosylation by inhibiting the transfer of UDP-N-acetylglucosamine (GlcNAc) to dolichol phosphate in the endoplasmic reticulum (ER) of eukaryotic cells, thus disrupting protein maturation [ 15, 16, 17 ].
How is tunicamycin used to treat gastric cancer?
Multidrug resistance remains a major obstacle to successful treatment for patients with gastric cancer (GC). Recently, glycosylation has been demonstrated to play a vital role in the acquisition of multidrug resistance. As a potent inhibitor of glycosylation, tunicamycin (Tu) has shown marked antitumor activities in various cancers.