Identification of galectin I and thioredoxin peroxidase II as two arsenic-binding proteins in Chinese hamster ovary cells.
نویسندگان
چکیده
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).
منابع مشابه
Proteomics Profiling of Chimeric-Truncated Tissue Plasminogen activator Producing- Chinese Hamster Ovary Cells Cultivated in a Chemically Defined Medium Supplemented with Protein Hydrolysates
Background: Culture media enrichment through the addition of protein hydrolysates is beneficial for achieving higher protein expression. Methods: In this study, designing the optimum mixture of four soy and casein-derived hydrolysates was successfully performed by design of experiment and specific productivity increased in all predicted combinations. Protein profile of recombinant CHO (rCHO) ce...
متن کاملP-234: Expression of Human Chorionic Gonadotropin (hCG) Hormone Using Chinese Hamster Ovary Cells
Background: Human chorionic gonadotropin (hCG) is a member of glycoprotein hormones family consist of two different non-covalently heterodimeric chains: alpha and beta subunits with 92 and 145 amino acids respectively. This hormone plays an important role in human reproduction and physiology especially for maintenance of the corpus luteum during the first months of pregnancy Materials and Metho...
متن کاملP-76: Stepwise Reduction of Fetal Bovine Serum Levels in Chinese Hamster OvaryCells -Expressing Human Chorionic Gonadotrophin- Culture
Background: The demand for producing recombinant therapeutic proteins by mammalian cell lines, such as Chinese hamster ovary (CHO) cells, continues to grow. Significant achievements in process optimization including development of cell culture strategies for largescale and cost-effective production have been made. Fetal bovine serum (FBS) is an often essential growth supplement and yet most cos...
متن کاملInterphase Death of Chinese Hamster Ovary Cells Exposed to Accelerated Heavy Ions
Introduction: Heavy ions are nucleus of elements of iron, argon, carbon and neon that all carry positive electrical charges. For these particles to be useful in radiotherapy they need to accelerated to high energy by more than thousand mega volts. Also the cosmic environment is considered to be a complicated mixture of highly energetic photons and heavy ions such as iron. Therefore, the health ...
متن کاملIntracellular Localization of FLAG-Peroxisomal Protein in Chinese Hamster Ovary (CHO) Cells
Epitope tagging is a method of expressing proteins whereby an epitope for a specific monoclonal antibody is fused to a target protein using recombinant DNA techniques. The aim of this study was to sub-clone the peroxisomal protein (PEP) cDNA into a mammalian expression vector leading to the formation of a chimeric PEP-cDNA containing the FLAG epitope. The FLAG-PEP recombinant cDNA was construc...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- The Biochemical journal
دوره 371 Pt 2 شماره
صفحات -
تاریخ انتشار 2003