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Uptake of copper from plasma proteins in cells where expression of CTR1 has been modulated

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Abstract

Plasma proteins rather than amino acid chelates are the direct sources of copper for mammalian cells. In continuing studies on the mechanisms by which albumin and transcuprein deliver copper and the potential involvement of CTR1, rates of uptake from these proteins and Cu–histidine were compared in cells with/without CTR1 knockdown or knockout. siRNA knocked down expression of CTR1 mRNA 60–85% in human mammary epithelial and hepatic cell models, but this had little or no effect on uptake of 1 μM Cu(II) attached to pure human albumin or alpha-2-macroglobulin. Mouse embryonic fibroblasts that did/did not express Ctr1 took up Cu(II) bound to albumin about as readily as from the histidine complex at physiological concentrations and by a single saturable process. Uptake from mouse albumin achieved a 2–4-fold higher Vmax (with a lower Km) than from heterologous human albumin. Maximum uptake rates from Cu(I)–histidine were >12-fold higher (with higher Km) than for Cu(II), suggesting mediation by a reductase. The presence of cell surface Cu(II) and Fe(III) reductase activity responding only slightly to dehydroascorbate was verified. Excess Fe(III) inhibited uptake from albumin–Cu(II). Ag(I) also inhibited, but kinetics were not or un-competitive. In general there was little difference in rates/kinetics of uptake in the Ctr1+/+ and −/− cells. Endocytosis was not involved. We conclude that plasma proteins deliver Cu(II) to homologous cells with greater efficiency than ionic copper at physiological concentrations, probably through the mediation of a Steap Cu(II)-reductase, and confirm the existence of an additional copper uptake system in mammalian cells.

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References

  • Abada P, Howell SB (2010) Regulation of cisplatin cytotoxicity by Cu influx transporters. Metal-Based Drugs 1–9 (article ID 317581)

  • Arredondo M, Munoz P, Mura CV, Nunez MT (2003) DMT1, a physiologically relevant Cu+1 transporter of intestinal cells. Am J Physiol (Cell Physiol) 284:C1525–C1530

    CAS  Google Scholar 

  • Bertinato J, Swist E, Plouffe LJ et al (2008) Ctr2 is partially localized to the plasma membrane and stimulates copper uptake in COS-7 cells. Biochem J 409:731–740

    Article  PubMed  CAS  Google Scholar 

  • Blair BG, Larson CA, Adams PL et al (2011) Copper transporter 2 regulates endocytosis and controls tumor growth and sensitivity to cisplatin in vivo. Mol Pharmacol 79:157–166

    Article  PubMed  CAS  Google Scholar 

  • Breslow E (1964) Comparison of cupric ion-binding sites in myoglobin derivatives and serum albumin. J Biol Chem 239:3252–3259

    PubMed  CAS  Google Scholar 

  • Cabrera A, Alonzo E, Chu Y-L et al (2008) Copper binding components of blood plasma and organs, and their responses to influx of large doses of 65Cu, in the mouse. Biometals 21:525–543

    Article  PubMed  CAS  Google Scholar 

  • Campbell CH, Brown RH, Linder MC (1981) Circulating ceruloplasmin is an important source of copper for normal and malignant cells. Bioch Biophys Acta 678:27–38

    Article  CAS  Google Scholar 

  • Festa RA, Thiele DJ (2011) Copper: an essential metal in biology. Curr Biol 21:R877–R883

    Article  PubMed  CAS  Google Scholar 

  • Gercken B, Barnes RM (1991) Determination of lead and other trace element species in blood by size exclusion chromatography and inductively coupled plasma/mass spectrometry. Anal Chem 63:283–287

    Google Scholar 

  • Gray LA, Kidane TZ, Nguyen A et al (2009) Multiple copper proteins and ferro-oxidases in mouse and human plasma. Biochem J 419:237–245

    Article  PubMed  CAS  Google Scholar 

  • Knopfel M, Solioz M (2002) Characterization of a cytochrome b558 ferric/cupric reductase from rabbit duodenal brush border membranes. Biochem Biophys Res Comm 291:220–225

    Article  PubMed  Google Scholar 

  • Knutson MD (2007) Steap proteins: implications for iron and copper metabolism. Nutr Rev 65:335–340

    Article  PubMed  Google Scholar 

  • Kuo Y-M, Zhou B, Cosco D et al (2001) The copper transporter CTR1 provides an essential function in mammalian embryonic development. Proc Natl Acad Sci USA 98:6836–6841

    Article  PubMed  CAS  Google Scholar 

  • Lau S, Sarkar B (1971) Ternary coordination complex between human serum albumin, copper(II) and l-histidine. J Biol Chem 246:5938–5943

    PubMed  CAS  Google Scholar 

  • Lee SH, Lancey R, Montaser A et al (1993) Ceruloplasmin and copper transport during the latter part of gestation in the rat. Proc Soc Exp Biol Med 203:428–439

    PubMed  CAS  Google Scholar 

  • Lee J, Prohaska JR, Thiele DJ (2001) Essential role for mammalian copper transporter Ctr1 in copper homeostasis and embryonic development. Proc Natl Acad Sci USA 98:6842–6847

    Article  PubMed  CAS  Google Scholar 

  • Lee J, Pena MMO, Nose Y et al (2002a) Biochemical characterization of the human copper transporter Ctr1. J Biol Chem 277:4380–4387

    Article  PubMed  CAS  Google Scholar 

  • Lee J, Petris MJ, Thiele DJ (2002b) Characterization of mouse embryonic cells deficient in the Ctr1 high affinity copper transporter: identification of a CTR1-independent copper transport system. J Biol Chem 277:40253–40259

    Article  PubMed  CAS  Google Scholar 

  • Linder MC (1991) Biochemistry of copper. Plenum Publishing, New York

    Google Scholar 

  • Linder MC (2002) Biochemistry and molecular biology of copper in mammals. In: Massoro EJ (ed) Handbook of copper pharmacology and toxicology. Humana Press, Totowa

    Google Scholar 

  • Linder MC (2010) Nutritional biochemistry of copper, with emphasis on the perinatal period. In: Avigliano L, Rossi L (eds) Biochemical aspects of human nutrition. Research Signpost, Trivandrum

    Google Scholar 

  • Linder MC, Zerounian NR, Moriya M et al (2003) Iron and copper homeostasis and intestinal absorption using the CaCO2 cell model. Biometals 16:145–160

    Article  PubMed  CAS  Google Scholar 

  • Liu NM, Lo LSL, Askary SH et al (2007) Transcuprein is a macroglobulin regulated by copper and iron availability. J Nutr Biochem 18:597–608

    Article  PubMed  Google Scholar 

  • Lutsenko S (2010) Human copper homeostasis: a network of interconnected pathways. Curr Opin Chem Biol 14:211–217

    Article  PubMed  CAS  Google Scholar 

  • Masuoka J, Hegenauer J, Van Dyke BR et al (1993) Intrinsic stoichiometric equilibrium constants for the binding of zinc(II) and copper(II) to the high affinity site of serum albumin. J Biol Chem 268:21533–21537

    PubMed  CAS  Google Scholar 

  • Mestek O, Kominkova J, Koplik R, Zima T, Miskusova M, Stern P (2002) Speciation of Cu, Se, Zn and Fe in blood serum of hemodialysed patients. Sb Lek 103:23–27

    Google Scholar 

  • Moriya M, Ho Y-H, Grana A et al (2008) Copper is taken up efficiently from albumin and alpha-2-macroglobulin by cultured human cells by more than one mechanism. Am J Physiol (Cell Physiol) 295:708–721

    Article  Google Scholar 

  • Musci G, Fraterrigo T, Calabrese L et al (1999) On the lability and functional significance of the type 1 copper pool in ceruloplasmin. J Biol Inorg Chem 4:441–446

    Article  PubMed  CAS  Google Scholar 

  • Ohgami RS, Campagne DR, McDonald A et al (2006) The Steap proteins are metalloreductases. Blood 108:1388–1394

    Article  PubMed  CAS  Google Scholar 

  • Olusanya O, Andrews PD, Swedlow JR et al (2001) Phosphorylation of threonine 156 of the mu2 subunit of the AP2 complex is essential for endocytosis in vitro and in vivo 11(11):896–900

  • Owen CA Jr (1971) Metabolism of copper 67 by the copper deficient rat. Am J Physiol 221:1722–1727

    PubMed  CAS  Google Scholar 

  • Rees EM, Thiele DJ (2007) Identification of a vacuole-associated metalloreductase and its role in Ctr2-mediated intracellular copper mobilization. J Biol Chem 282:21629–21638

    Article  PubMed  CAS  Google Scholar 

  • Scott KC, Turnlund JR (1994) Compartmental model of copper metabolism in adult men. J Nutr Biochem 5:342–350

    Article  CAS  Google Scholar 

  • Shum SCK, Houk RS (1993) Elemental speciation by anion exchange and size exclusion chromatography with detection by inductively coupled plasma mass spectrometry with direct injection nebulization. Anal Chem 65:2972–2976

    Article  CAS  Google Scholar 

  • Turner JR, Tartakoff AM (1989) The response of the golgi complex to microtubule alterations: The roles of metabolic energy and membrane traffic in golgi complex organization. J Cell Biol 109:2081–2088

    Article  PubMed  CAS  Google Scholar 

  • Turnlund JR, Keyes WR, Peiffer GL et al (1998) Copper absorption, excretion, and retention by young men consuming low dietary copper determined by using the stable isotope 65Cu. Am J Clin Nutr 67:1219–1225

    PubMed  CAS  Google Scholar 

  • van den Berghe PV, Folmer DE, Malingre HE et al (2007) Human copper transporter 2 is localized in late endosomes and lysosomes and facilitates cellular copper uptake. Biochem J 407:49–59

    Article  PubMed  Google Scholar 

  • Vargas JD, Herpers B, McKie AT et al (2003) Stromal cell-derived receptor 2 and cytochrome b561 are functional ferric reductases. Biochim Biophys Acta 1651:116–123

    PubMed  CAS  Google Scholar 

  • Weiss KC, Linder MC (1985) Copper transport in rats, involving a new plasma protein. Am J Physiol Endocrinol Metab 249:E77–E88

    CAS  Google Scholar 

  • Wirth PL, Linder MC (1985) Distribution of copper among components of human serum. J Natl Cancer Inst 75:277–284

    Google Scholar 

  • Wyman S, Simpson RJ, McKie AT et al (2008) Dcytb (Cybrd1) functions as both a ferric and a cupric reductase in vitro. FEBS Lett 582:1901–1906

    Article  PubMed  CAS  Google Scholar 

  • Zhou B, Gitschier J (1997) hCTR1: a human gene for copper uptake identified by complementation in yeast. Proc Natl Acad Sci USA 94:7481–7486

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We gratefully acknowledge receipt of mouse embryonic fibroblasts that do and do not express Ctr1 from Dennis Thiele at Duke University. Supported in part by Public Health Service Grant No. RO1 HD46949 for the research, and IR24CA86307 to the Mallinkrodt Institute of Radiology (MIR) at Washington University, St. Louis, subsidizing 64Cu production. Ramin Farhad, Eric Russo, and Kyoung Jin Lee were supported by an Howard Hughes Medical Institute grant for the CSUF-HHMI Undergraduate Research Scholars Program. No conflicts of interest leading to financial or other gain are connected with the research reported here.

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  1. Department of Chemistry and Biochemistry, California State University, Fullerton, CA, 92834-6866, USA

    Theodros Z. Kidane, Ramin Farhad, Kyoung Jin Lee, Abraham Santos, Eric Russo & Maria C. Linder

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  1. Theodros Z. Kidane
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  2. Ramin Farhad
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  3. Kyoung Jin Lee
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  4. Abraham Santos
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  5. Eric Russo
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  6. Maria C. Linder
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Correspondence to Maria C. Linder.

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Kidane, T.Z., Farhad, R., Lee, K.J. et al. Uptake of copper from plasma proteins in cells where expression of CTR1 has been modulated. Biometals 25, 697–709 (2012). https://doi.org/10.1007/s10534-012-9528-8

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  • DOI: https://doi.org/10.1007/s10534-012-9528-8

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