Calreticulin transacetylase mediates the acetylation of nitric oxide synthase by polyphenolic acetate

Autor: Elwin Verheij, Seema Bansal, Ramesh C. Rastogi, Yogesh K Tyagi, Virinder S. Parmar, Ajit Kumar, Prija Ponnan, Marco Gaspari, Hanumantharao G. Raj, Giovanni Cuda
Přispěvatelé: TNO Kwaliteit van Leven
Rok vydání: 2008
Předmět:
coumarin derivative
Calcium-Binding Protein
Vitamin D-Dependent
Placenta
Lysine
Polyphenolic acetates
enzyme purification
Nitric Oxide Synthase Type I
Reductase
Acetates
Applied Microbiology and Biotechnology
Biochemistry
calreticulin
chemistry.chemical_compound
Pregnancy
Tandem Mass Spectrometry
acyltransferase
binding affinity
Nanotechnology
Enzyme activity
polyphenol derivative
mass spectrometry
biology
nitric oxide synthase
article
protein domain
Acetylation
General Medicine
Nitric oxide synthase
Calbindin 2
calreticulin transacetylase
Transacetylase
Female
Biotechnology
Binding sites
PDZ domain
Protein domain
Molecular Sequence Data
Amino-Acid N-Acetyltransferase
Bioengineering
Protein acetylation
polyphenol acetate
In Vitro Techniques
7
8 diacetoxy 4 methylcoumarin
Nitric oxide
S100 Calcium Binding Protein G
Phenols
Acetyltransferases
reduced nicotinamide adenine dinucleotide phosphate
Humans
controlled study
human
Amino Acid Sequence
Molecular Biology
Flavonoids
lysine
catalysis
Proteins
Polyphenols
human tissue
Cell membranes
chemistry
biology.protein
Peptides
Calreticulin
Chromatography
Liquid
Zdroj: Applied Biochemistry and Biotechnology, 1, 144, 37-45
ISSN: 0273-2289
Popis: Our earlier investigations identified acetoxy drug: protein transacetylase (TAase), a unique enzyme in the endoplasmic reticulum (ER) catalyzing the transfer of acetyl groups from polyphenolic acetates (PA) to certain functional proteins. Recently we have established the identity of TAase with ER protein calreticulin (CR) and subsequently transacetylase function of CR was termed calreticulin transacetylase (CRTAase). CRTAase was purified and characterized from human placenta. CRTAase catalyzed the acetylation of a receptor protein nNOS, by a model PA 7, 8-diacetoxy-4-methylcoumarin (DAMC), which was visually confirmed by using antiacetyl lysine. The aim of this report was to provide tacit proof by providing mass spectrometry evidence for CRTAase catalyzed acetylation of purified nNOS by DAMC. For this purpose, purified nNOS was incubated with DAMC and CRTAase, the modified nNOS was analyzed by nanoscale LC-MS/MS, which recorded 11 distinct peptides with a significant score as acetylated on lysine residues. The distribution was in order: lysines-24, -33, -38, -131, and -229 of the PDZ domain, Lys-245 of the oxygenase domain, Lys-754 and -856 of FMN binding domain, Lys-989 of connecting domain and Lys-1300, -1321, and -1371 of the NADPH-binding domain were acetylated. The results documented in this paper highlighted for the first time modification of nNOS by way of acetylation. Our earlier work recorded the profound activation of platelet NADPH cytochrome P-450 reductase and the acetylation of the reductase protein by DAMC, which also remarkably enhanced intracellular levels of nitric oxide. The results reported here coupled with the aforementioned previous observations strongly implicate the possible role of the acetylation of the reductase domain of nitric oxide synthase (NOS) in the NOS activation. In addition, the acetylation of nNOS can be expected to potentiate the interaction with CR, eventually leading to the augmented catalytic activity of NOS and expression of the related biological effects. © 2007 Humana Press Inc.
Databáze: OpenAIRE
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