Protein Acyltransferase Function of Purified Calreticulin: The Exclusive Role of P-Domain in Mediating Protein Acylation Utilizing Acyloxycoumarins and Acetyl CoA as the Acyl Group Donors

Singh, Prabhjot; Ponnan, Prija; Priya, Nivedita; Tyagi, T.K.; Gaspari, Marco; Krishnan, Shibu; Cuda, Giovanni; Joshi, Pallavi; Gambhir, Jasvinder K.; Sharma, Sunil; Prasad, Ashok K.; Saso, Luciano; Rastogi, R. C.; Parmar, Virinder S.; Raj, Hanumantharao G.

doi:10.2174/092986611794927938

Cited by 8 publications

(9 citation statements)

References 25 publications

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“…Interestingly, we have observed that the full length hCR and the recombinant P-domain possess CRTAase activity, while the N- and C-domains had no CRTAase activity. P-domain catalyzed acetylation of recombinant GST from Schistosoma japonicum (rGST), which have remarkable sequence and structural identity with GST3-3 (an isoform of mu class of rat GST), was analyzed by using the acetylated lysine antibody and by nanoscale LC-MS/MS, which identified the modification of several lysine residues on rGST [62]. …”

Section: Novel Function Of Calreticulin: Protein Acetyltransferasementioning

confidence: 99%

“…Also, acetyl CoA is a rather large molecule, with a molecular mass of 810 Da whereas the molecular mass of DAMC is 276 Da.The lysine acetylation is a result of a small alteration of the amino acid side chain in the local secondary structure, it could prevent the long β -mercaptoethylamine and pantothenate moiety of acetyl CoA from obtaining efficient entry into and exit from the ER lumen. It is worth noting that P-domain catalyzed acetylation of rGST as revealed by LC-MS/MS spectra resulted in the modification of several lysine residues in common, when either DAMC or acetyl CoA was used as the acetyl group donor [62]. …”

Section: Calreticulin As An Acetyl Coa-linked Protein Acetyl Transmentioning

confidence: 99%

“…The lysine acetylated sites were located by LC-MS/MS analysis [14, 62]. In case of S. japonicum GST Lys-9 and Lys-40 forming α -helices, Lys-27 forming β -turns, lied in the N-terminal α / β domain of the protein, Lys-78 was the only acetylated lysine residue in the linker region of S. japonicum GST and Lys-87, -119, -125, -131, -181, -194, and -191 ( α -helix) and Lys-113 ( β -turn) lied in C-terminal α -domain.…”

Section: Computational Investigations On Acetylated Gst the Produmentioning

confidence: 99%

“…We have extended the investigations to explore the ability to transfer propionyl group to the receptor protein rGST by rhCRTAase utilizing propoxycoumarin as the propionyl group donor. CRTAase was reported to utilize acetyl CoA as the acetyl group donor [62]. …”

Section: Points Of Similarities and Dissimilarities Between Crtaasmentioning

confidence: 99%

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Comparison of Protein Acetyltransferase Action of CRTAase with the Prototypes of HAT

Ponnan

Kumar

Singh

et al. 2014

The Scientific World Journal

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Our laboratory is credited for the discovery of enzymatic acetylation of protein, a phenomenon unknown till we identified an enzyme termed acetoxy drug: protein transacetylase (TAase), catalyzing the transfer of acetyl group from polyphenolic acetates to receptor proteins (RP). Later, TAase was identified as calreticulin (CR), an endoplasmic reticulum luminal protein. CR was termed calreticulin transacetylase (CRTAase). Our persistent study revealed that CR like other families of histone acetyltransferases (HATs) such as p300, Rtt109, PCAF, and ESA1, undergoes autoacetylation. The autoacetylated CR was characterized as a stable intermediate in CRTAase catalyzed protein acetylation, and similar was the case with ESA1. The autoacetylation of CR like that of HATs was found to enhance protein-protein interaction. CR like HAT-1, CBP, and p300 mediated the acylation of RP utilizing acetyl CoA and propionyl CoA as the substrates. The similarities between CRTAase and HATs in mediating protein acylation are highlighted in this review.

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Section: Novel Function Of Calreticulin: Protein Acetyltransferasementioning

confidence: 99%

Section: Calreticulin As An Acetyl Coa-linked Protein Acetyl Transmentioning

confidence: 99%

Section: Computational Investigations On Acetylated Gst the Produmentioning

confidence: 99%

Section: Points Of Similarities and Dissimilarities Between Crtaasmentioning

confidence: 99%

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Comparison of Protein Acetyltransferase Action of CRTAase with the Prototypes of HAT

Ponnan

Kumar

Singh

et al. 2014

The Scientific World Journal

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“…Interestingly, calreticulin, an ER luminal protein that is mostly known as a Ca +2 -binding chaperone, appears to have acetyl-CoA:lysine acetyltransferase activity in vitro [40]. Whether calreticulin retains acetyltransferase activity in vivo remains to be determined.…”

Section: A Novel Form Of Post-translational Regulation In the Ermentioning

confidence: 99%

Lysine acetylation in the lumen of the ER: A novel and essential function under the control of the UPR

Pehar

Puglielli

2013

Biochimica et Biophysica Acta (BBA) - Molecular Cell Research

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The Nε-amino group of lysine residues can be transiently modified by the addition of an acetyl group. Recognized functions of Nε-lysine acetylation include regulation of activity, molecular stabilization and conformational assembly of a protein. For more than forty years lysine acetylation was thought to occur only in the cytosol and nucleus. Targets included cytoskeletal-associated proteins as well as transcription factors, histone proteins and proteins involved in DNA recombination and repair. However, in 2007 we reported that a type I membrane protein involved in the pathogenesis of Alzheimer’s disease was transiently acetylated on the ε amino group of seven lysine residues while transiting along the secretory pathway. Surprisingly, the acetylation occurred in the lumen of the endoplasmic reticulum (ER) forcing us to reconsider old paradigms. Indeed, if lysine acetylation can occur in the lumen of the ER, then all the essential biochemical elements of the reaction must be available in the lumen of the organelle. Follow-up studies revealed the existence of ER-based acetyl-CoA:lysine acetyltransferases as well as a membrane transporter that translocates acetyl-CoA from the cytosol into the ER lumen. Large-scale proteomics showed that the list of substrates of the ER-based acetylation machinery includes both transiting and resident proteins. Finally, genetic studies revealed that this machinery is tightly linked to human diseases. Here, we describe these exciting findings as well as recent biochemical and cellular advances, and discuss possible impact on both human physiology and pathology.

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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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Protein Acyltransferase Function of Purified Calreticulin: The Exclusive Role of P-Domain in Mediating Protein Acylation Utilizing Acyloxycoumarins and Acetyl CoA as the Acyl Group Donors

Cited by 8 publications

References 25 publications

Comparison of Protein Acetyltransferase Action of CRTAase with the Prototypes of HAT

Comparison of Protein Acetyltransferase Action of CRTAase with the Prototypes of HAT

Lysine acetylation in the lumen of the ER: A novel and essential function under the control of the UPR

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

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