Digestive Diseases and Sciences

, Volume 52, Issue 8, pp 1979–1986

Proteomic Analysis of Protein Plugs: Causative Agent of Symptoms in Patients with Choledochal Cyst

  • Kenitiro Kaneko
  • Hisami Ando
  • Takahiko Seo
  • Yasuyuki Ono
  • Takahisa Tainaka
  • Wataru Sumida
Original Paper

Abstract

Symptoms of choledochal cysts are caused by protein plugs. We performed proteomic analysis of protein plugs to elucidate formation mechanism. Protein plugs were obtained from three pediatric patients with choledochal cyst. Proteins were separated using sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Gel bands common to the samples were excised for mass spectrometry. Mass spectra were compared with the NCBI database for protein identification. Gel bands of protein plug samples were predominant at 14 kilodaltons (kDa), followed by 29 kDa. Four other thin bands were common to the plug samples. Four bands (including 14 and 29 kDa) were identified as lithostathine, and one band as serum albumin. Plugs consisted mostly of lithostathine, a protein secreted by pancreatic acinar cells into pancreatic juice. The mechanism involves trypsinogen and lithostathine regurgitating into the cyst through an aberrant union of pancreaticobiliary ducts. Activated trypsin cleaves soluble lithostathine into insoluble forms that aggregate to form plugs.

Keywords

Choledochal cyst Pancreaticobiliary maljunction Protein plugs Mechanism of plug formation Proteomics 

References

  1. 1.
    Todani T, Watanabe Y, Naruse M, Tabuchi K, Okajima K (1977) Congenital bile duct cysts: classification, operative procedures, and review of thirty-seven cases including cancer arising from choledochal cyst. Am J Surg 134:263–269PubMedCrossRefGoogle Scholar
  2. 2.
    Todani T (1997) Congenital choledochal dilatation: classification, clinical features, and long-term results. J Hepatobil Pancreat Surg 4:276–282CrossRefGoogle Scholar
  3. 3.
    Visser BC, Suh I, Way LW, Kang SM (2004) Congenital choledochal cysts in adults. Arch Surg 139:855–862PubMedCrossRefGoogle Scholar
  4. 4.
    Japanese Study Group on Pancreaticobiliary Maljunction (JSPBM), Committee of JSPBM for Diagnostic Criteria (1994) Diagnostic criteria of pancreaticobiliary maljunction. J Hepatobil Pancreat Surg 1:219–221CrossRefGoogle Scholar
  5. 5.
    Kaneko K, Ando H, Ito T, Watanabe Y, Seo T, Harada T, Ito F (1997) Protein plugs cause symptoms in patients with choledochal cysts. Am J Gastroenterol 92:1018–1021PubMedGoogle Scholar
  6. 6.
    Kaneko K, Ando H, Ito T, Kasai K, Watanabe Y, Seo T (1996) Increased cell proliferation and transforming growth factor-alpha (TGF alpha) in the gall-bladder epithelium of patients with pancreaticobiliary maljunction. Pathol Int 46:253–260PubMedCrossRefGoogle Scholar
  7. 7.
    Kaneko K, Ando H, Seo T, Ono Y, Ochiai K, Ogura Y (2005) Bile infection contributes to intrahepatic calculi formation after excision of choledochal cysts. Pediatr Surg Int 21:8–11PubMedCrossRefGoogle Scholar
  8. 8.
    Maki T (1966) Pathogenesis of calcium bilirubinate stone: role of E. coli, b-glucuronidase and coagulation by inorganic ions, polyelectrolytes and agitation. Ann Surg 164:90–100PubMedCrossRefGoogle Scholar
  9. 9.
    Ando H, Ito T, Kaneko K, Seo T, Ito F (1996) Intrahepatic bile duct stenosis causing intrahepatic calculi formation following excision of a choledochal cyst. J Am Coll Surg 183:56–60PubMedGoogle Scholar
  10. 10.
    Uno K, Tsuchida Y, Kawarasaki H, Ohmiya H, Honna T (1996) Development of intrahepatic cholelithiasis long after primary excision of choledochal cysts. J Am Coll Surg 183:583–588PubMedGoogle Scholar
  11. 11.
    Watanabe Y, Toki A, Todani T (1999) Bile duct cancer developed after cyst excision for choledochal cyst. J Hepatobil Pancreat Surg 6:207–212CrossRefGoogle Scholar
  12. 12.
    Kobayashi S, Asano T, Yamasaki M, Kenmochi T, Nakagohri T, Ochiai T (1999) Risk of bile duct carcinogenesis after excision of extrahepatic bile ducts in pancreaticobiliary maljunction. Surgery 126:939–944PubMedCrossRefGoogle Scholar
  13. 13.
    Goto N, Yasuda I, Uematsu T, Kanemura N, Takai S, Ando K, Kato T, Osada S, Takao H, Saji S, Shimokawa K, Moriwaki H (2001) Intrahepatic cholangiocarcinoma arising 10 years after the excision of congenital extrahepatic biliary dilation. J Gastroenterol 36:856–862PubMedCrossRefGoogle Scholar
  14. 14.
    Tsuchida A, Kasuya K, Endo M, Saito H, Inoue K, Nagae I, Aoki T, Koyanagi Y (2003) High risk of bile duct carcinogenesis after primary resection of a congenital biliary dilatation. Oncol Rep 10:1183–1187PubMedGoogle Scholar
  15. 15.
    Ando H, Ito T, Watanabe Y, Seo T, Kaneko K, Nagaya M (1995) Spontaneous perforation of choledochal cyst. J Am Coll Surg 181:125–128PubMedGoogle Scholar
  16. 16.
    Ando H, Ito T, Nagaya M, Watanabe Y, Seo T, Kaneko K (1995) Pancreaticobiliary maljunction without choledochal cysts in infants and children: clinical features and surgical therapy. J Pediatr Surg 30:1658–1662PubMedCrossRefGoogle Scholar
  17. 17.
    Shevchenko A, Wilm M, Vorm O, Mann M (1996) Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal Chem 68:850–858PubMedCrossRefGoogle Scholar
  18. 18.
    Giorgi D, Bernard JP, Rouquier S, Iovanna J, Sarles H, Dagorn JC (1989) Secretory pancreatic stone protein messenger RNA. Nucleotide sequence and expression in chronic calcifying pancreatitis. J Clin Invest 84:100–106PubMedCrossRefGoogle Scholar
  19. 19.
    Watanabe T, Yonekura H, Terazono K, Yamamoto H, Okamoto H (1990) Complete nucleotide sequence of human reg gene and its expression in normal and tumoral tissues: the reg protien, pancreatic stone protein, and pancreatic thread protein are one and the same product of the gene. J Biol Chem 265:7432–7439PubMedGoogle Scholar
  20. 20.
    Patard L, Lallemand JY, Stoven V (2003) An insight into the role of human pancreatic lithostathine. JOP 4:92–103PubMedGoogle Scholar
  21. 21.
    Provansal-Cheylan M, Mariani A, Bernard JP, Sarles H, Dupuy P (1989) Pancreatic stone protein: quantification in pancreatic juice by enzyme-linked immunosorbent assay and comparison with other methods. Pancreas 4:680–689PubMedCrossRefGoogle Scholar
  22. 22.
    Schmiegel W, Burchert M, Kalthoff H, Roeder C, Butzow G, Grimm H, Kremer B, Soehendra N, Schreiber HW, Thiele HG, Greten H (1990) Immunochemical characterization and quantitative distribution of pancreatic stone protein in sera and pancreatic secretions in pancreatic disorders. Gastroenterology 99:1421–1430PubMedGoogle Scholar
  23. 23.
    Lee BI, Mustafi D, Cho W, Nakagawa Y (2003) Characterization of calcium binding properties of lithostathine. J Biol Inorg Chem 8:341–347PubMedGoogle Scholar
  24. 24.
    Gregoire C, Marco S, Thimonier J, Duplan L, Laurine E, Chauvin JP, Michel B, Peyrot V, Verdier JM (2001) Three-dimensional structure of the lithostathine protofibril, a protein involved in Alzheimer's disease. EMBO J 20:3313–3321PubMedCrossRefGoogle Scholar
  25. 25.
    Graf R, Schiesser M, Scheele GA, Marquardt K, Frick TW, Ammann RW, Bimmler D (2001) A family of 16-kDa pancreatic secretory stress proteins form highly organized fibrillar structures upon tryptic activation. J Biol Chem 276:21028–21038PubMedCrossRefGoogle Scholar
  26. 26.
    Iovanna JL, Dagorn JC (2005) The multifunctional family of secreted proteins containing a C-type lectin-like domain linked to a short N-terminal peptide. Biochim Biophys Acta 1723:8–18PubMedGoogle Scholar
  27. 27.
    Guy O, Robles-Diaz G, Adrich Z, Sahel J, Sarles H (1983) Protein content of precipitates present in pancreatic juice of alcoholic subjects and patients with chronic calcifying pancreatitis. Gastroenterology 84:102–107PubMedGoogle Scholar
  28. 28.
    Multigner L, Daudon M, Montalto G, De Caro A, Etienne JP, Sarles H (1986) Radiolucent pancreatic stones. N Engl J Med 314:248PubMedGoogle Scholar
  29. 29.
    Mariani A, Bernard JP, Provansal-Cheylan M, Nitsche S, Sarles H (1991) Differnces of pancreatic stone morphology and content in patients with pancreatic lithiasis. Dig Dis Sci 36:1509–1516CrossRefPubMedGoogle Scholar
  30. 30.
    Ozturk M, de la Monte SM, Gross J, Wands JR (1989) Elevated leveld of an exocrine pancreatic secretory protein in Alzheimer disease brain. Proc Natl Acad Sci USA 86:419–423PubMedCrossRefGoogle Scholar
  31. 31.
    Laurine E, Gregoire C, Fandrich M, Engemann S, Marchal S, Thion L, Mohr M, Monsarrat B, Michel B, Dobson CM, Wanker E, Erard M, Verdier JM (2003) Lithostathine quadruple-helical filaments form proteinase K-resistant deposits in Creutzfeldt-Jakob disease. J Bio Chem 51:51770–51778CrossRefGoogle Scholar
  32. 32.
    Duplan L, Michel B, Boucraut J, Barthellemy S, Desplat-Jego S, Marin V, Gambarelli D, Bernard D, Berthezene P, Alescio-Lautier B, Verdier JM (2001) Lithostathine and pancreatitis-associated protein are involved in the very early stages of Alzheimer's disease. Neurobiol Aging 22:79–88PubMedCrossRefGoogle Scholar
  33. 33.
    Kaneko K, Ando H, Watanabe Y, Seo T, Harada T (2000) Pathologic changes in the common bile duct of an experimental model with pancreaticobiliary maljunction. Pediatr Surg Int 16:26–28PubMedCrossRefGoogle Scholar
  34. 34.
    Kristiansen TZ, Bunkenborg J, Gronborg M, Molina H, Thuluvath PJ, Argani P, Goggins MG, Maitra A, Pandey A (2004) A proteomic analysis of human bile. Mol Cell Proteomics 3:715–728PubMedCrossRefGoogle Scholar
  35. 35.
    Zhou H, Chen B, Li RX, Sheng QH, Li SJ, Zhang L, Li L, Xia QC, Wang HY, Zeng R (2005) Large-scale identification of human biliary proteins from a cholesterol stone patient using a proteomic approach. Rapid Commun Mass Spectrom 19;3569–3578PubMedCrossRefGoogle Scholar
  36. 36.
    Grønborg M, Bunkenborg J, Kristiansen TZ, Jensen ON, Yeo CJ, Hruban RH, Maitra A, Goggins MG, Pandey A (2004) Comprehensive proteomic analysis of human pancreatic juice. J Proteome Res 3:1042–1055PubMedCrossRefGoogle Scholar
  37. 37.
    He XM, Carter DC (1992) Atomic structure and chemistry of human serum albumin. Nature 358:209–215PubMedCrossRefGoogle Scholar
  38. 38.
    Smits ME, Groen AK, Mok KS, van Marle J, Tytgat GN, Huibregtse K (1997) Analysis of occluded pancreatic stents and juices in patients with chronic pancreatitis. Gastrointest Endosc 45:52–58PubMedCrossRefGoogle Scholar
  39. 39.
    Farnbacher MJ, Voll RE, Faissner R, Wehler M, Hahn EG, Lohr M, Schneider HT (2005) Composition of clogging material in pancreatic endoprostheses. Gastrointest Endosc 61:862–866PubMedCrossRefGoogle Scholar
  40. 40.
    Ochiai K, Kaneko K, Kitagawa M, Ando H, Hayakawa T (2004) Activated pancreatic enzyme and pancreatic stone protein (PSP/reg) in bile of patients with pancreaticobiliary maljunction/choledochal cyst. Dig Dis Sci 49:1953–1956PubMedCrossRefGoogle Scholar
  41. 41.
    Bensimon G, Lacomblez L, Meininger V (1994) A controlled trial of riluzole in amyotrophic lateral sclerosis. N Engl J Med 330:585–591PubMedCrossRefGoogle Scholar
  42. 42.
    Heiser V, Engemann S, Brocker W, Dunkel I, Boeddrich A, Waelter S, Nordhoff E, Lurz R, Schugardt N, Rautenberg S, Herhaus C, Barnickel G, Bottcher H, Lehrach H, Wanker EE (2002) Identification of benzothiazoles as potential polyglutamine aggregation inhibitors of Huntington's disease by using an automated filter retardation assay. Proc Natl Acad Sci USA 99:16400–16406PubMedCrossRefGoogle Scholar
  43. 43.
    Huntington Study Group (2003) Dosage effects of riluzole in Huntington's disease. A multicenter placebo-controlled study. Neurology 61:1551–1556Google Scholar
  44. 44.
    Tsuchida A, Itoi T, Kasuya K, Endo M, Katsumata K, Aoki T, Suzuki M, Aoki T (2005) Inhibitory effect of meloxicam, a cyclooxygenase-2 inhibitor, on N-nitrosobis (2-oxopropyl) amine induced biliary carcinogenesis in Syrian hamsters. Carcinogenesis 26:1922–1928PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Kenitiro Kaneko
    • 1
  • Hisami Ando
    • 1
  • Takahiko Seo
    • 1
  • Yasuyuki Ono
    • 1
  • Takahisa Tainaka
    • 1
  • Wataru Sumida
    • 1
  1. 1.Department of Pediatric SurgeryNagoya University Graduate School of MedicineShowa-kuJapan

Personalised recommendations