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Identification of galectin I and thioredoxin peroxidase II as two arsenic-binding proteins in Chinese hamster ovary cells.
Biochem J. 2003 Apr 15; 371(Pt 2):495-503.BJ

Abstract

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 20 mM 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).

Authors+Show Affiliations

Department of Life Science, National Central University, Chung-Li, Taoyuan, Taiwan 32054, Republic of China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

12519079

Citation

Chang, Kwang Ning, et al. "Identification of Galectin I and Thioredoxin Peroxidase II as Two Arsenic-binding Proteins in Chinese Hamster Ovary Cells." The Biochemical Journal, vol. 371, no. Pt 2, 2003, pp. 495-503.
Chang KN, Lee TC, Tam MF, et al. Identification of galectin I and thioredoxin peroxidase II as two arsenic-binding proteins in Chinese hamster ovary cells. Biochem J. 2003;371(Pt 2):495-503.
Chang, K. N., Lee, T. C., Tam, M. F., Chen, Y. C., Lee, L. W., Lee, S. Y., Lin, P. J., & Huang, R. N. (2003). Identification of galectin I and thioredoxin peroxidase II as two arsenic-binding proteins in Chinese hamster ovary cells. The Biochemical Journal, 371(Pt 2), 495-503.
Chang KN, et al. Identification of Galectin I and Thioredoxin Peroxidase II as Two Arsenic-binding Proteins in Chinese Hamster Ovary Cells. Biochem J. 2003 Apr 15;371(Pt 2):495-503. PubMed PMID: 12519079.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Identification of galectin I and thioredoxin peroxidase II as two arsenic-binding proteins in Chinese hamster ovary cells. AU - Chang,Kwang Ning, AU - Lee,Te Chang, AU - Tam,Ming F, AU - Chen,Yi Chin, AU - Lee,Li Wen, AU - Lee,Shin Ying, AU - Lin,Pei Jung, AU - Huang,Rong Nan, PY - 2003/01/09/accepted PY - 2002/12/18/revised PY - 2002/08/29/received PY - 2003/1/10/pubmed PY - 2003/7/4/medline PY - 2003/1/10/entrez SP - 495 EP - 503 JF - The Biochemical journal JO - Biochem J VL - 371 IS - Pt 2 N2 - 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 20 mM 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). SN - 0264-6021 UR - https://www.unboundmedicine.com/medline/citation/12519079/Identification_of_galectin_I_and_thioredoxin_peroxidase_II_as_two_arsenic_binding_proteins_in_Chinese_hamster_ovary_cells_ L2 - https://portlandpress.com/biochemj/article-lookup/doi/10.1042/BJ20021354 DB - PRIME DP - Unbound Medicine ER -