Cellular and Molecular Life Sciences

, Volume 68, Issue 23, pp 3919–3931

Clusterin facilitates in vivo clearance of extracellular misfolded proteins

  • Amy R. Wyatt
  • Justin J. Yerbury
  • Paula Berghofer
  • Ivan Greguric
  • Andrew Katsifis
  • Christopher M. Dobson
  • Mark R. Wilson
Research article

Abstract

The extracellular deposition of misfolded proteins is a characteristic of many debilitating age-related disorders. However, little is known about the specific mechanisms that act to suppress this process in vivo. Clusterin (CLU) is an extracellular chaperone that forms stable and soluble complexes with misfolded client proteins. Here we explore the fate of complexes formed between CLU and misfolded proteins both in vitro and in a living organism. We show that proteins injected into rats are cleared more rapidly from circulation when complexed with CLU as a result of their more efficient localization to the liver and that this clearance is delayed by pre-injection with the scavenger receptor inhibitor fucoidan. The CLU–client complexes were found to bind preferentially, in a fucoidan-inhibitable manner, to human peripheral blood monocytes and isolated rat hepatocytes and in the latter cell type were internalized and targeted to lysosomes for degradation. The data suggest, therefore, that CLU plays a key role in an extracellular proteostasis system that recognizes, keeps soluble, and then rapidly mediates the disposal of misfolded proteins.

Keywords

Clusterin Extracellular chaperone Misfolded protein Receptor-mediated endocytosis Clearance 

Supplementary material

18_2011_684_MOESM1_ESM.tif (8.6 mb)
Supplementary Fig. 1 Time-dependent accumulation of the injected dose in the thyroid of Sprague–Dawley rats after injection with 123I-labeled HMW CLU-stressed protein complexes or control proteins. a Pseudocolor images of the distribution of radioactivity in the head and upper body of a rat (orientation indicated by the labels head and abdomen) injected with 123I-HMW CLU–FGN via the tail vein. The times indicated are p.i. Progressively higher levels of radioactivity are indicated by the color gradient starting at blue and moving through green, yellow, red and finally white. The images shown are representative of three different experiments. The arrow indicates the position of the thyroid gland. b Panels shows the proportion of the injected dose per gram of thyroid up to 30 min after the animals were injected with 123I-labeled HMW CLU–client complexes or uncomplexed control protein as indicated. Note, the mass of the rat thyroid is less than 1 g, so the percentage values plotted can exceed 100%. Data points represent means (n = 4 ± standard error) and are corrected for any radioactivity remaining in the tail
18_2011_684_MOESM2_ESM.tif (1.2 mb)
Supplementary Fig. 2 The binding of biotinylated HMW CLU–client protein complexes and control proteins to rat liver cells, assessed by flow cytometry. Enriched preparations of hepatocytes or non-parenchymal liver cells were incubated with biotinylated a FGN, residual soluble heated FGN (FGN#) or HMW CLU–FGN, b GST, residual soluble heated GST (GST#) or HMW CLU-GST, or c CLU or residual soluble heated CLU (CLU#), and then SA-Alexa Fluor® 488. The results shown are the geometric mean of the Alexa Fluor® 488 fluorescence intensity in AU (n = 3 ± standard error)

References

  1. 1.
    Dobson CM (2003) Protein folding and misfolding. Nature 426:884–890PubMedCrossRefGoogle Scholar
  2. 2.
    Ben-Zvi A, Miller EA, Morimoto RI (2009) Collapse of proteostasis represents an early molecular event in Caenorhabditis elegans aging. Proc Natl Acad Sci USA 106:14914–14919PubMedCrossRefGoogle Scholar
  3. 3.
    Wyatt AR, Yerbury JJ, Poon S, Wilson MR (2009) Therapeutic targets in extracellular protein deposition diseases. Curr Med Chem 16:2855–2866PubMedCrossRefGoogle Scholar
  4. 4.
    Yerbury JJ, Stewart EM, Wyatt AR, Wilson MR (2005) Quality control of protein folding in extracellular space. EMBO Rep 6:1131–1136PubMedCrossRefGoogle Scholar
  5. 5.
    Hartl FU, Hayer-Hartl M (2002) Molecular chaperones in the cytosol: from nascent chain to folded protein. Science 295:1852–1857PubMedCrossRefGoogle Scholar
  6. 6.
    Iwao Y, Anraku M, Yamasaki K, Kragh-Hansen U, Kawai K et al (2006) Oxidation of Arg-410 promotes the elimination of human serum albumin. Biochim Biophys Acta 1764:743–749PubMedGoogle Scholar
  7. 7.
    Margineanu I, Ghetie V (1981) A selective model of plasma protein catabolism. J Theor Biol 90:101–110PubMedCrossRefGoogle Scholar
  8. 8.
    Wilson MR, Yerbury JJ, Poon S (2008) Potential roles of abundant extracellular chaperones in the control of amyloid formation and toxicity. Mol BioSys 4:42–52CrossRefGoogle Scholar
  9. 9.
    Humphreys DT, Carver JA, Easterbrook-Smith SB, Wilson MR (1999) Clusterin has chaperone-like activity similar to that of small heat shock proteins. J Biol Chem 274:6875–6881PubMedCrossRefGoogle Scholar
  10. 10.
    French K, Yerbury JJ, Wilson MR (2008) Protease activation of alpha-2-macroglobulin modulates a chaperone-like broad specificity. Biochemistry 47:1176–1185PubMedCrossRefGoogle Scholar
  11. 11.
    Yerbury JJ, Rybchyn MS, Easterbrook-Smith SB, Henriques C, Wilson MR (2005) The acute phase protein haptoglobin is a mammalian extracellular chaperone with an action similar to clusterin. Biochemistry 44:10914–10925PubMedCrossRefGoogle Scholar
  12. 12.
    Matsudomi N, Kanda Y, Yoshika Y, Moriwaki H (2004) Ability of alpha-S-casein to suppress the heat aggregation of ovotransferrin. J Agric Food Chem 52:4882–4886PubMedCrossRefGoogle Scholar
  13. 13.
    Zhang X, Fu X, Zhang H, Liu C, Jiao W et al (2005) Chaperone-like activity of beta-casein. Int J Biochem Cell Biol 37:1232–1240PubMedCrossRefGoogle Scholar
  14. 14.
    Wyatt AR, Yerbury JJ, Wilson MR (2009) Structural characterization of clusterin–client protein complexes. J Biol Chem 284:21920–21927PubMedCrossRefGoogle Scholar
  15. 15.
    Bell RD, Sagare AP, Friedman AE, Bedi GS, Holtzman DM et al (2007) Transport pathways for clearance of human Alzheimer’s amyloid beta-peptide and apolipoproteins E and J in the mouse central nervous system. J Cereb Blood Flow Metab 27:909–918PubMedGoogle Scholar
  16. 16.
    Rosenberg ME, Girton R, Finkel D, Chmielewski D, Barrie A et al (2002) Apolipoprotein J/clusterin prevents progressive glomerulopathy of aging. Mol Cell Biol 22:1893–1902PubMedCrossRefGoogle Scholar
  17. 17.
    Wilson MR, Yerbury JJ, Poon S (2008) Extracellular chaperones and amyloids, In: Asea A, Brown I (eds) Heat Shock proteins and the brain: implications for neurodegenerative diseases and neuroprotection. Springer, Netherlands, pp 283–315Google Scholar
  18. 18.
    Wyatt AR, Wilson MR (2010) Identification of human plasma proteins as major clients for the extracellular chaperone clusterin. J Biol Chem 285:3532–3539PubMedCrossRefGoogle Scholar
  19. 19.
    Lee HB, Blaufox MD (1985) Blood volume in the rat. J Nuc Med 26:72–76Google Scholar
  20. 20.
    Zlokovic BV, Martel CL, Matsubara E, McComb JG, Zheng G et al (1996) Glycoprotein 330 megalin: probable role in receptor-mediated transport of apolipoprotein J alone and in a complex with Alzheimer disease amyloid β at the blood–brain and blood–cerebrospinal fluid barriers. Proc Natl Acad Sci USA 93:4229–4234PubMedCrossRefGoogle Scholar
  21. 21.
    Kounnas MZ, Loukinova EB, Steffansson S, Harmony JAK, Brewer BH et al (1995) Identification of glycoprotein 330 as an endocytic receptor for apolipoprotein J/clusterin. J Biol Chem 270:13070–13075PubMedCrossRefGoogle Scholar
  22. 22.
    Lundgren S, Carling T, Hjälm G, Juhlin C, Rastad J et al (1997) Tissue distribution of human gp330/megalin, a putative Ca2+ sensing protein. J Histochem Cytochem 45:383–392PubMedCrossRefGoogle Scholar
  23. 23.
    Pluddemann A, Neyen C, Gordon S (2007) Macrophage scavenger receptors and host-derived ligands. Methods 43:207–217PubMedCrossRefGoogle Scholar
  24. 24.
    Gowen BB, Borg TK, Ghaffar A, Mayer EP (2001) The collagenous domain of class A scavenger receptors is involved in macrophage adhesion to collagens. J Leukoc Biol 69:575–582PubMedGoogle Scholar
  25. 25.
    Horiuchi S, Sakamoto Y, Sakai M (2003) Scavenger receptors for oxidized and glycated proteins. Amino Acids 25:283–292PubMedCrossRefGoogle Scholar
  26. 26.
    Herczenik E, Gebbink MFBG (2008) Molecular and cellular aspects of protein misfolding and disease. FASEB J 22:2115–2133PubMedCrossRefGoogle Scholar
  27. 27.
    Berteau O, Mulloy B (2003) Sulfated fucans, fresh perspectives: structures, functions, and biological properties of sulfated fucans and an overview of enzymes active toward this class of polysaccharide. Glycobiology 13:29R–40RPubMedCrossRefGoogle Scholar
  28. 28.
    Draude G, Hrboticky N, Lorenz RL (1999) The expression of the lectin-like oxidized low-density lipoprotein receptor (LOX-1) on human vascular smooth muscle cells and monocytes and its down-regulation by lovastatin. Biochem Pharmacol 57:383–386PubMedCrossRefGoogle Scholar
  29. 29.
    Sakamoto H, Aikawa M, Hill CC, Weiss D, Robert Taylor W et al (2001) Biomechanical strain induces class A scavenger receptor expression in human monocyte/macrophages and THP-1 cells. Circulation 104:109–114PubMedCrossRefGoogle Scholar
  30. 30.
    Platt N, Suzuki H, Kurihara Y, Kodama T, Gordon S (1996) Role for the class A macrophage scavenger receptor in the phagocytosis of apoptotic thymocytes in vitro. Proc Natl Acad Sci USA 93:12456–12460PubMedCrossRefGoogle Scholar
  31. 31.
    Oka K, Sawamura T, Kikuta K, Itokawa S, Kume N et al (1998) Lectin-like oxidized low-density lipoprotein receptor 1 mediates phagocytosis of aged/apoptotic cells in endothelial cells. Proc Natl Acad Sci USA 95:9535–9540PubMedCrossRefGoogle Scholar
  32. 32.
    Berwin B, Delneste Y, Lovingood RV, Post SR, Pizzo SV (2004) SREC-I, a type F scavenger receptor, is an endocytic receptor for calreticulin. J Biol Chem 279:51250–51257PubMedCrossRefGoogle Scholar
  33. 33.
    Granert C, Raud J, Xie X, Lindquist L, Lindbom L (1994) Inhibition of leukocyte rolling with polysaccharide fucoidin prevents pleocytosis in experimental meningitis in the rabbit. J Clin Invest 93:929–936PubMedCrossRefGoogle Scholar
  34. 34.
    Ley K, Linnemann G, Meinen M, Stoolman LM, Graehtgens P (1993) Fucoidin, but not yeast polyphosphomannan PPME, inhibits leukocyte rolling in venules of the rat mesentery. Blood 81:177–185PubMedGoogle Scholar
  35. 35.
    Johnson JD, Hess KL, Cook-Mills JM (2003) CD44, alpha 4 integrin, and fucoidin receptor-mediated phagocytosis of apoptotic leukocytes. J Leukoc Biol 74:810–820PubMedCrossRefGoogle Scholar
  36. 36.
    Platt N, Gordon S (1998) Scavenger receptors: diverse activities and promiscuous binding of polyanionic ligands. Chem Biol 5:R193–R203PubMedCrossRefGoogle Scholar
  37. 37.
    Poon S, Rybchyn MS, Easterbrook-Smith SB, Carver JA, Wilson MR (2000) Clusterin is an ATP-independent chaperone with a very broad substrate specificity that stabilizes stressed proteins in a folding-competent state. Biochemistry 39:15953–15960PubMedCrossRefGoogle Scholar
  38. 38.
    Cardona-Sanclemente LE, Born GVR (1995) Effect of inhibition of nitric oxide synthesis on the uptake of LDL and fibrinogen by arterial walls and other organs of the rat. Br J Pharmacol 114:1490–1494PubMedGoogle Scholar
  39. 39.
    Bartl MM, Luckenbach T, Bergner O, Ullrich O, Koch-Brandt C (2001) Multiple receptors mediate apoJ-dependent clearance of cellular debris into nonprofessional phagocytes. Exp Cell Res 271:130–141PubMedCrossRefGoogle Scholar
  40. 40.
    Aarseth P, Klug D (1972) Dehydration-induced reductions in total blood volume and in pulmonary blood volume in rats. Acta Physiol Scand 85:277–282PubMedCrossRefGoogle Scholar
  41. 41.
    Rigotti A, Miettinen HE, Kreiger M (2003) The role of high-density lipoprotein receptor SR-BI in the lipid metabolism of endocrine and other tissues. Endocr Rev 24:357–387PubMedCrossRefGoogle Scholar
  42. 42.
    Yuasa H, Watanabe J (2003) Are novel scavenger-like receptors involved in the hepatic clearance of heparin? Drug Metab Pharmacokinet 18:273–286PubMedCrossRefGoogle Scholar
  43. 43.
    Mahon MG, Linstedt KA, Hermann M, Nimpf J, Schneider WJ (1999) Multiple involvement of clusterin in chicken ovarian follicle development. J Biol Chem 274:4036–4044PubMedCrossRefGoogle Scholar
  44. 44.
    Bajari TM, Strasser V, Nimpf J, Schneider WJ (2003) A model for modulation of leptin activity by association with clusterin. FASEB J 17:1505–1507PubMedGoogle Scholar
  45. 45.
    Galantai R, Modos K, Fidy J, Kolev K, Machovich R (2006) Structural basis of the cofactor function of denatured albumin in plasminogen activation by tissue-type plasminogen activator. Biochem Biophys Res Comm 341:736–741PubMedCrossRefGoogle Scholar
  46. 46.
    Kranenburg OBB, Kroon-Batenburg LMJ, Reijerkerk A, Wu YP et al (2002) Tissue-type plasminogen activator is a multiligand cross-β structure receptor. Curr Biol 12:1833–1839PubMedCrossRefGoogle Scholar
  47. 47.
    Gidalevitza T, Kikisa EA, Morimoto RI (2010) A cellular perspective on conformational disease: the role of genetic background and proteostasis networks. Curr Opin Struct Biol 20:23–32CrossRefGoogle Scholar

Copyright information

© Springer Basel AG 2011

Authors and Affiliations

  • Amy R. Wyatt
    • 1
    • 2
  • Justin J. Yerbury
    • 1
    • 2
  • Paula Berghofer
    • 3
  • Ivan Greguric
    • 3
  • Andrew Katsifis
    • 3
  • Christopher M. Dobson
    • 4
  • Mark R. Wilson
    • 1
    • 2
  1. 1.School of Biological SciencesUniversity of WollongongWollongongAustralia
  2. 2.Illawarra Health and Medical Research Institute, University of WollongongWollongongAustralia
  3. 3.Radiopharmaceutical Research InstituteAustralian Nuclear Science and Technology OrganisationLucas HeightsAustralia
  4. 4.Department of ChemistryUniversity of CambridgeCambridgeUK

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