Calreticulin transacetylase: A novel enzyme-mediated protein acetylation by acetoxy derivatives of 3-alkyl-4-methylcoumarins

Jalal, Sarah; Chand, Karam; Kathuria, Abha; Singh, Prabhjot; Priya, Nivedita; Gupta, Bhavna; Raj, Hanumantharao G.; Sharma, Sunil

doi:10.1016/j.bioorg.2011.10.005

Cited by 9 publications

(4 citation statements)

References 21 publications

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“…[ 10 ] DAMC was synthesized according to previous work. [ 11 ] DPeMC was a novel ligand synthesized by our laboratory. [ 7 ] Both structures were confirmed through 1 H nuclear magnetic resonance (NMR), infrared (IR) spectroscopy, and liquid chromatography mass spectrometry (LCMS).…”

Section: Methodsmentioning

confidence: 99%

Spectroscopic studies of 7,8‐diacetoxy‐4‐methylcoumarin and 7,8‐dipentynoyl‐4‐methylcoumarin binding with calreticulin

et al. 2022

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Calreticulin (CRT) is a protein found mainly in the endoplasmic reticulum (ER) that maintains calcium levels and controls protein folding, but has recently been found at the cell surface, cytoplasm, and in the extracellular matrix. CRT participates in multiple physiological processes such as gene expression, the immune response, and cancer. Calreticulin has been shown to autoacetylate with the binding of preferred ligand 7,8‐diacetoxy‐4‐methylcoumarin (DAMC). This project aims to develop a chemical biology approach to investigate importance of CRT acylating abilities on its nonendoplasmic reticulum functions by targeting the downstream substrates of CRT acetylation. Our goal was to use CRT to transfer a pentynoyl tag (using a novel ligand, DPeMC) to its substrates, which can then be used as a handle for protein identification. Molecular modelling using available data in the literature was used to approximate the binding interface between CRT and the acylation ligands. Molecular Operating Environment (MOE) software was used to perform sequence alignment, simulated annealing, positional refinement, and blind docking of acylated coumarins with the CRT model. Docking studies pointed to the P domain as the most thermodynamically and sterically favourable region for acylated coumarin binding with tryptophan residue 200 within the active site on CRT. Absorption and fluorescence spectra of all coumarin compounds in ethanol:PBS (1:9 v/v) solution were investigated. Stern–Volmer quenching constant (KSV), binding constant (K), and number of binding sites (n) of each coumarin compound with CRT was determined. Our studies demonstrated that acyl coumarin compounds bind to CRT using a dynamic quenching mechanism, bind to a single binding site on the P domain, and that the protein–ligand interaction is spontaneous and exothermic.

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Section: Methodsmentioning

confidence: 99%

Spectroscopic studies of 7,8‐diacetoxy‐4‐methylcoumarin and 7,8‐dipentynoyl‐4‐methylcoumarin binding with calreticulin

et al. 2022

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“…After separation by flash chromatography on silica gel (pentane/ethyl acetate from 100/0 to 95/5), 4a was isolated as two diastereomers in 23% and 22% yields, respectively (79 mg, 0.25 mmol, 45% overall yield). Ethyl 2-acetylpentanoate (5b) 45 Diethylzinc (1.12 mL, 1 M in heptane, 2 equiv) was added under argon to a room-temperature solution of 2-bromo-acrylic acid ethyl ester (100 mg, 0.56 mmol, 1 equiv) and acetyltrimethylsilane (89 µL, 0.62 mmol, 1.1 equiv) in nondegassed dichloromethane (2.8 mL). After 18 h at room temperature, the reaction was quenched with saturated NH 4 Cl.…”

Section: A C C E P T E D Accepted Manuscriptmentioning

confidence: 99%

Theoretical investigation of the reaction of dialkylzincs with α-alkoxycarbonyl radicals. Evaluation of α-bromoacrylates as radical acceptors in radical-polar crossover processes

et al. 2015

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“…7,8-Diacetoxy-4-methylcoumarin (1). 4-Methyl-2-oxo-2Hchromene-7,8-diyl diacetate (DAMC) was synthesized using a previously reported procedure (Jalal et al, 2012).…”

Section: Synthesis and Crystallizationmentioning

confidence: 99%

Crystal structures of 4-methyl-2-oxo-2H-chromene-7,8-diyl diacetate and 4-methyl-2-oxo-2H-chromene-7,8-diyl bis(pent-4-ynoate)

et al. 2016

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Calreticulin transacetylase: A novel enzyme-mediated protein acetylation by acetoxy derivatives of 3-alkyl-4-methylcoumarins

Cited by 9 publications

References 21 publications

Spectroscopic studies of 7,8‐diacetoxy‐4‐methylcoumarin and 7,8‐dipentynoyl‐4‐methylcoumarin binding with calreticulin

Spectroscopic studies of 7,8‐diacetoxy‐4‐methylcoumarin and 7,8‐dipentynoyl‐4‐methylcoumarin binding with calreticulin

Theoretical investigation of the reaction of dialkylzincs with α-alkoxycarbonyl radicals. Evaluation of α-bromoacrylates as radical acceptors in radical-polar crossover processes

Crystal structures of 4-methyl-2-oxo-2H-chromene-7,8-diyl diacetate and 4-methyl-2-oxo-2H-chromene-7,8-diyl bis(pent-4-ynoate)

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