Surface heat shock protein 90 serves as a potential receptor for calcium oxalate crystal on apical membrane of renal tubular epithelial cells

  • Kedsarin Fong-ngern
  • Kanyarat Sueksakit
  • Visith ThongboonkerdEmail author
Original Paper


Adhesion of calcium oxalate monohydrate (COM) crystals on renal tubular epithelial cells is a crucial step in kidney stone formation. Finding potential crystal receptors on the apical membrane of the cells may lead to a novel approach to prevent kidney stone disease. Our previous study identified a large number of crystal-binding proteins on the apical membrane of MDCK cells. However, their functional role as potential crystal receptors had not been validated. The present study aimed to address the potential role of heat shock protein 90 (HSP90) as a COM crystal receptor. The apical membrane was isolated from polarized MDCK cells by the peeling method and recovered proteins were incubated with COM crystals. Western blot analysis confirmed the presence of HSP90 in the apical membrane and the crystal-bound fraction. Immunofluorescence staining without permeabilization and laser-scanning confocal microscopy confirmed the surface HSP90 expression on the apical membrane of the intact cells. Crystal adhesion assay showed that blocking surface HSP90 by specific anti-HSP90 antibody and knockdown of HSP90 by small interfering RNA (siRNA) dramatically reduced crystal binding on the apical surface of MDCK cells (by approximately 1/2 and 2/3, respectively). Additionally, crystal internalization assay revealed the presence of HSP90 on the membrane of endocytic vesicle containing the internalized COM crystal. Moreover, pretreatment of MDCK cells with anti-HSP90 antibody significantly reduced crystal internalization (by approximately 1/3). Taken together, our data indicate that HSP90 serves as a potential receptor for COM crystals on the apical membrane of renal tubular epithelial cells and is involved in endocytosis/internalization of the crystals into the cells.


Apical membrane Calcium oxalate Crystal adhesion Crystal binding Crystal receptor 



This study was supported by Mahidol University research grant, Office of the Higher Education Commission and Mahidol University under the National Research Universities Initiative, and the Thailand Research Fund (RTA5680004). VT is supported by “Chalermphrakiat”, whereas KF is supported by Faculty of Medicine Siriraj Hospital.


  1. 1.
    Lieske JC, Deganello S, Toback FG (1999) Nephron 81(Suppl 1):8–17PubMedGoogle Scholar
  2. 2.
    Thongboonkerd V (2008) Contrib Nephrol 160:142–158CrossRefPubMedGoogle Scholar
  3. 3.
    Verkoelen CF, Verhulst A (2007) Kidney Int 72:13–18CrossRefPubMedGoogle Scholar
  4. 4.
    Kumar V, Farell G, Deganello S, Lieske JC (2003) J Am Soc Nephrol 14:289–297CrossRefPubMedGoogle Scholar
  5. 5.
    Sorokina EA, Kleinman JG (1999) J Biol Chem 274:27491–27496CrossRefPubMedGoogle Scholar
  6. 6.
    Verkoelen CF, van der Boom BG, Romijn JC (2000) Kidney Int 58:1045–1054CrossRefPubMedGoogle Scholar
  7. 7.
    Sorokina EA, Wesson JA, Kleinman JG (2004) J Am Soc Nephrol 15:2057–2065CrossRefPubMedGoogle Scholar
  8. 8.
    Take M, Tsutsui J, Obama H, Ozawa M, Nakayama T, Maruyama I, Arima T, Muramatsu T (1994) J Biochem 116:1063–1068PubMedGoogle Scholar
  9. 9.
    Verkoelen CF, van der Boom BG, Kok DJ, Romijn JC (2000) Kidney Int 57:1072–1082CrossRefPubMedGoogle Scholar
  10. 10.
    Kohjimoto Y, Ebisuno S, Tamura M, Ohkawa T (1996) Scanning Microsc 10:459–468PubMedGoogle Scholar
  11. 11.
    Fong-ngern K, Peerapen P, Sinchaikul S, Chen ST, Thongboonkerd V (2011) J Proteome Res 10:4463–4477CrossRefPubMedGoogle Scholar
  12. 12.
    Jin S, Song YC, Emili A, Sherman PM, Chan VL (2003) Cell Microbiol 5:165–174CrossRefPubMedGoogle Scholar
  13. 13.
    Reyes-Del Valle J, Chavez-Salinas S, Medina F, Del Angel RM (2005) J Virol 79:4557–4567CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Fullekrug J, Simons K (2004) Ann N Y Acad Sci 1014:164–169CrossRefPubMedGoogle Scholar
  15. 15.
    Rindler MJ, Chuman LM, Shaffer L, Saier MH Jr (1979) J Cell Biol 81:635–648CrossRefPubMedGoogle Scholar
  16. 16.
    Saier MH Jr (1981) Am J Physiol 240:C106–C109PubMedGoogle Scholar
  17. 17.
    Thongboonkerd V, Semangoen T, Chutipongtanate S (2006) Clin Chim Acta 367:120–131CrossRefPubMedGoogle Scholar
  18. 18.
    Chaiyarit S, Mungdee S, Thongboonkerd V (2010) Anal Methods 2:1536–1541CrossRefGoogle Scholar
  19. 19.
    Fong-ngern K, Chiangjong W, Thongboonkerd V (2009) Anal Biochem 395:25–32CrossRefPubMedGoogle Scholar
  20. 20.
    Burns JR, Finlayson B (1980) Invest Urol 18:167–169PubMedGoogle Scholar
  21. 21.
    Chaiyarit S, Thongboonkerd V (2012) J Proteome Res 11:3269–3280CrossRefPubMedGoogle Scholar
  22. 22.
    Meder D, Shevchenko A, Simons K, Fullekrug J (2005) J Cell Biol 168:303–313CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Delacour D, Cramm-Behrens CI, Drobecq H, Le Bivic A, Naim HY, Jacob R (2006) Curr Biol 16:408–414CrossRefPubMedGoogle Scholar
  24. 24.
    Tsutsumi S, Neckers L (2007) Cancer Sci 98:1536–1539CrossRefPubMedGoogle Scholar
  25. 25.
    Lieske JC, Swift H, Martin T, Patterson B, Toback FG (1994) Proc Natl Acad Sci 91:6987–6991CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Kanlaya R, Sintiprungrat K, Chaiyarit S, Thongboonkerd V (2013) Cell Biochem Biophys 67:1171–1179CrossRefPubMedGoogle Scholar
  27. 27.
    Cid C, Regidor I, Poveda PD, Alcazar A (2009) Cell Stress Chaperones 14:321–327CrossRefPubMedGoogle Scholar
  28. 28.
    Camins A, Diez-Fernandez C, Prieto P (1999) Toxicol In Vitro 13:437–443CrossRefPubMedGoogle Scholar
  29. 29.
    Slimane TA, Trugnan G, van Ijzendoorn SC, Hoekstra D (2003) Mol Biol Cell 14:611–624CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Somji S, Ann SM, Garrett SH, Gurel V, Todd JH, Sens DA (2002) Toxicol Lett 133:241–254CrossRefPubMedGoogle Scholar
  31. 31.
    Lin TW, Lo CW, Lai SY, Fan RJ, Lo CJ, Chou YM, Thiruvengadam R, Wang AH, Wang MY (2007) J Virol 81:8730–8741CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Koul HK, Menon M, Chaturvedi LS, Koul S, Sekhon A, Bhandari A, Huang M (2002) J Biol Chem 277:36845–36852CrossRefPubMedGoogle Scholar
  33. 33.
    Peerapen P, Thongboonkerd V (2013) Sci Rep 3:1041CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Evan AP, Lingeman JE, Coe FL, Parks JH, Bledsoe SB, Shao Y, Sommer AJ, Paterson RF, Kuo RL, Grynpas M (2003) J Clin Invest 111:607–616CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Kumar V, Farell G, Yu S, Harrington S, Fitzpatrick L, Rzewuska E, Miller VM, Lieske JC (2006) J Investig Med 54:412–424CrossRefPubMedGoogle Scholar
  36. 36.
    Evan AP, Coe FL, Lingeman JE, Shao Y, Sommer AJ, Bledsoe SB, Anderson JC, Worcester EM (2007) Anat Rec (Hoboken) 290:1315–1323CrossRefGoogle Scholar
  37. 37.
    Singhto N, Sintiprungrat K, Thongboonkerd V (2013) J Proteome Res 12:3561–3572CrossRefPubMedGoogle Scholar

Copyright information

© SBIC 2016

Authors and Affiliations

  • Kedsarin Fong-ngern
    • 1
  • Kanyarat Sueksakit
    • 1
  • Visith Thongboonkerd
    • 1
    Email author
  1. 1.Medical Proteomics Unit, Office for Research and Development, Faculty of MedicineSiriraj Hospital, Center for Research in Complex Systems Science, Mahidol UniversityBangkokThailand

Personalised recommendations