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Identification of arsenite-and arsenic diglutathione-binding proteins in human hepatocarcinoma cells

Abstract

It is generally accepted that trivalent arsenicals are more toxic than the corresponding pentavalent arsenicals, since trivalent arsenicals bind the thiol groups of biomolecules, leading to a deterioration in cellular functions. In the present study, we prepared three different arsenic-bound sepharoses and investigated the binding of hepatic cytosolic proteins to pentavalent, trivalent, and glutathione-conjugated trivalent arsenicals. SDS-PAGE showed no proteins bound to pentavalent arsenic specifically. In contrast, we found a number of proteins that have specific and high affinity for trivalent arsenic. Two of those proteins were identified: protein disulfide isomerase-related protein 5 (PDSIRP5) and peroxiredoxin 1/enhancer protein (PRX1/EP). These proteins have vicinal cysteines, as previously reported. In contrast, one of the prominent proteins that did not bind to trivalent arsenic was identified as calreticulin precursor. Although there are 3 cysteines in calreticulin precursor, two of the cysteines are spaced more than 25 amino acids apart. Five synthetic peptides containing 2 vicinal cysteines were prepared to study whether they would inhibit the binding of PDSIRP5, PRX1/EP, and other arsenic-binding proteins to trivalent arsenicals. Only two of the five peptides effectively inhibited binding, suggesting that other amino acids besides the 2 vicinal cysteines may modulate the affinity of cysteine-rich proteins for  More>>
Publication Date:
Jan 15, 2010
Product Type:
Journal Article
Resource Relation:
Journal Name: Toxicology and Applied Pharmacology; Journal Volume: 242; Journal Issue: 2; Other Information: DOI: 10.1016/j.taap.2009.10.013; PII: S0041-008X(09)00451-7; Copyright (c) 2009 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.
Subject:
60 APPLIED LIFE SCIENCES; ARSENIC; BILE; CYSTEINE; DETOXIFICATION; GLUTATHIONE; HEPATOMAS; LIVER; MEN; METABOLISM; TOXICITY; AMINO ACIDS; ANIMALS; BIOLOGICAL MATERIALS; BODY; BODY FLUIDS; CARBOXYLIC ACIDS; CARCINOMAS; DIGESTIVE SYSTEM; DISEASES; DRUGS; ELEMENTS; GLANDS; MALES; MAMMALS; MAN; MATERIALS; NEOPLASMS; ORGANIC ACIDS; ORGANIC COMPOUNDS; ORGANIC SULFUR COMPOUNDS; ORGANS; PEPTIDES; POLYPEPTIDES; PRIMATES; PROTEINS; RADIOPROTECTIVE SUBSTANCES; RESPONSE MODIFYING FACTORS; SEMIMETALS; THIOLS; VERTEBRATES
OSTI ID:
21344832
Country of Origin:
United States
Language:
English
Other Identifying Numbers:
Journal ID: ISSN 0041-008X; TXAPA9; TRN: US10R1673075734
Availability:
Available from http://dx.doi.org/10.1016/j.taap.2009.10.013
Submitting Site:
INIS
Size:
page(s) 119-125
Announcement Date:
Oct 26, 2010

Citation Formats

Mizumura, Ayano, Watanabe, Takayuki, Kobayashi, Yayoi, Environmental Health Sciences Division, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki 305-8506 (Japan)], Hirano, Seishiro, and Research Center for Environmental Risk, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki 305-8506 (Japan)]. Identification of arsenite-and arsenic diglutathione-binding proteins in human hepatocarcinoma cells. United States: N. p., 2010. Web. doi:10.1016/j.taap.2009.10.013.
Mizumura, Ayano, Watanabe, Takayuki, Kobayashi, Yayoi, Environmental Health Sciences Division, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki 305-8506 (Japan)], Hirano, Seishiro, & Research Center for Environmental Risk, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki 305-8506 (Japan)]. Identification of arsenite-and arsenic diglutathione-binding proteins in human hepatocarcinoma cells. United States. doi:10.1016/j.taap.2009.10.013.
Mizumura, Ayano, Watanabe, Takayuki, Kobayashi, Yayoi, Environmental Health Sciences Division, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki 305-8506 (Japan)], Hirano, Seishiro, and Research Center for Environmental Risk, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki 305-8506 (Japan)]. 2010. "Identification of arsenite-and arsenic diglutathione-binding proteins in human hepatocarcinoma cells." United States. doi:10.1016/j.taap.2009.10.013. https://www.osti.gov/servlets/purl/10.1016/j.taap.2009.10.013.
@misc{etde_21344832,
title = {Identification of arsenite-and arsenic diglutathione-binding proteins in human hepatocarcinoma cells}
author = {Mizumura, Ayano, Watanabe, Takayuki, Kobayashi, Yayoi, Environmental Health Sciences Division, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki 305-8506 (Japan)], Hirano, Seishiro, and Research Center for Environmental Risk, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki 305-8506 (Japan)]}
abstractNote = {It is generally accepted that trivalent arsenicals are more toxic than the corresponding pentavalent arsenicals, since trivalent arsenicals bind the thiol groups of biomolecules, leading to a deterioration in cellular functions. In the present study, we prepared three different arsenic-bound sepharoses and investigated the binding of hepatic cytosolic proteins to pentavalent, trivalent, and glutathione-conjugated trivalent arsenicals. SDS-PAGE showed no proteins bound to pentavalent arsenic specifically. In contrast, we found a number of proteins that have specific and high affinity for trivalent arsenic. Two of those proteins were identified: protein disulfide isomerase-related protein 5 (PDSIRP5) and peroxiredoxin 1/enhancer protein (PRX1/EP). These proteins have vicinal cysteines, as previously reported. In contrast, one of the prominent proteins that did not bind to trivalent arsenic was identified as calreticulin precursor. Although there are 3 cysteines in calreticulin precursor, two of the cysteines are spaced more than 25 amino acids apart. Five synthetic peptides containing 2 vicinal cysteines were prepared to study whether they would inhibit the binding of PDSIRP5, PRX1/EP, and other arsenic-binding proteins to trivalent arsenicals. Only two of the five peptides effectively inhibited binding, suggesting that other amino acids besides the 2 vicinal cysteines may modulate the affinity of cysteine-rich proteins for trivalent arsenicals. We further investigated hepatic cytosolic proteins that bound specifically to glutathione-conjugated trivalent arsenic, which is the most abundant form of arsenical in bile fluid. Four proteins that bound specifically to glutathione-conjugated trivalent arsenic were identified; interestingly, these proteins were different from the trivalent arsenic-binding proteins. These results suggest that although glutathione-conjugation is an important process in the metabolism, excretion, and detoxification of arsenicals, glutathione-conjugated arsenicals can still react with some proteins in hepatic cells.}
doi = {10.1016/j.taap.2009.10.013}
journal = {Toxicology and Applied Pharmacology}
issue = {2}
volume = {242}
place = {United States}
year = {2010}
month = {Jan}
}