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. doi: 10.1042/BJ20021354.

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).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Arsenic / metabolism*
  • Base Sequence
  • CHO Cells
  • Carrier Proteins / metabolism
  • Chromatography, High Pressure Liquid
  • Cricetinae
  • DNA Primers
  • DNA, Complementary / genetics
  • Galectin 1 / genetics
  • Galectin 1 / isolation & purification
  • Galectin 1 / metabolism*
  • Humans
  • Kinetics
  • Mice
  • Molecular Sequence Data
  • Neoplasm Proteins*
  • Peptide Fragments / chemistry
  • Peroxidases / genetics
  • Peroxidases / isolation & purification
  • Peroxidases / metabolism*
  • Peroxiredoxin III
  • Peroxiredoxins
  • Rats
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Alignment
  • Sequence Homology, Amino Acid

Substances

  • Carrier Proteins
  • DNA Primers
  • DNA, Complementary
  • Galectin 1
  • Neoplasm Proteins
  • Peptide Fragments
  • Prdx3 protein, mouse
  • Peroxidases
  • PRDX3 protein, human
  • Peroxiredoxin III
  • Peroxiredoxins
  • Arsenic

Associated data

  • GENBANK/AF221841