In this study, we report the identification of two arsenic-binding proteins from Chinese hamster ovary (CHO) cells. The crude extract derived from CHO and SA7 (arsenic-resistant CHO cells) was applied to a phenylarsine oxide–agarose affinity column, and after extensive washing, the absorbed proteins were eluted with buffers containing 20mM 2-mercaptoethanol (2-ME) or dithiothreitol (DTT). Three differentially expressed proteins, galectin 1 (Gal-1; in the 2-ME-eluted fraction from CHO cells), glutathione S-transferase P-form (GST-P) and thioredoxin peroxidase II (TPX-II), respectively in the 2-ME- and DTT-eluted fractions from SA7 cells, were identified by partial amino acid sequence analysis after separation by SDS/PAGE. The GST-P protein has been previously shown to facilitate the excretion of sodium arsenite [As(III)] from SA7 cells. TPX II was detected predominately in SA7 cells [routinely cultured in As(III)-containing medium], but not in CHO or SA7N (a revertant of SA7 cells cultured in regular medium) cells. In contrast, Gal-1 was specifically identified in CHO and SA7N cells, but not in SA7 cells. The preferential expression of Gal-1 in CHO cells and TPX-II in SA7 cells was further illustrated by quantitative PCR analysis. The binding of Gal-1 and TPX-II with As(III) was further verified by both co-immunoprecipitation and co-elution of Gal-1 and TPX-II with As(III). It is suggested that Gal-1 and TPX-II are two proteins that serve as high-affinity binding sites for As(III) and thus both may be involved in the biological action of As(III).
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April 2003
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Research Article|
April 15 2003
Identification of galectin I and thioredoxin peroxidase II as two arsenic-binding proteins in Chinese hamster ovary cells
Kwang Ning CHANG;
Kwang Ning CHANG
∗Department of Life Science, National Central University, Chung-Li, Taoyuan, Taiwan 32054, Republic of China
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T. Chang LEE;
T. Chang LEE
†Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan, 11529, Republic of China
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Ming F. TAM;
Ming F. TAM
‡Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan, 11529, Republic of China
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Y. Chin CHEN;
Y. Chin CHEN
∗Department of Life Science, National Central University, Chung-Li, Taoyuan, Taiwan 32054, Republic of China
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L. Wen LEE;
L. Wen LEE
∗Department of Life Science, National Central University, Chung-Li, Taoyuan, Taiwan 32054, Republic of China
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Shin Ying LEE;
Shin Ying LEE
∗Department of Life Science, National Central University, Chung-Li, Taoyuan, Taiwan 32054, Republic of China
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Pei Jung LIN;
Pei Jung LIN
∗Department of Life Science, National Central University, Chung-Li, Taoyuan, Taiwan 32054, Republic of China
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Rong Nan HUANG
Rong Nan HUANG
1
∗Department of Life Science, National Central University, Chung-Li, Taoyuan, Taiwan 32054, Republic of China
1To whom correspondence should be addressed (e-mail LSRONG@cc.ncu.edu.tw).
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Publisher: Portland Press Ltd
Received:
August 29 2002
Revision Received:
December 18 2002
Accepted:
January 09 2003
Accepted Manuscript online:
January 09 2003
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2003
2003
Biochem J (2003) 371 (2): 495–503.
Article history
Received:
August 29 2002
Revision Received:
December 18 2002
Accepted:
January 09 2003
Accepted Manuscript online:
January 09 2003
Citation
Kwang Ning CHANG, T. Chang LEE, Ming F. TAM, Y. Chin CHEN, L. Wen LEE, Shin Ying LEE, Pei Jung LIN, Rong Nan HUANG; Identification of galectin I and thioredoxin peroxidase II as two arsenic-binding proteins in Chinese hamster ovary cells. Biochem J 15 April 2003; 371 (2): 495–503. doi: https://doi.org/10.1042/bj20021354
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