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Bansal S Gaspari M Raj HG Kumar A Cuda G Verheij E Tyagi YK Ponnan P Rastogi RC Parmar VS 《Applied biochemistry and biotechnology》2008,144(1):37-45
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. 相似文献
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