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

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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.

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References

  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 134:263–269

    Article  PubMed  CAS  Google Scholar 

  2. Todani T (1997) Congenital choledochal dilatation: classification, clinical features, and long-term results. J Hepatobil Pancreat Surg 4:276–282

    Article  Google Scholar 

  3. Visser BC, Suh I, Way LW, Kang SM (2004) Congenital choledochal cysts in adults. Arch Surg 139:855–862

    Article  PubMed  Google Scholar 

  4. Japanese Study Group on Pancreaticobiliary Maljunction (JSPBM), Committee of JSPBM for Diagnostic Criteria (1994) Diagnostic criteria of pancreaticobiliary maljunction. J Hepatobil Pancreat Surg 1:219–221

    Article  Google Scholar 

  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 92:1018–1021

    PubMed  CAS  Google Scholar 

  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 46:253–260

    Article  PubMed  CAS  Google Scholar 

  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 21:8–11

    Article  PubMed  Google Scholar 

  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 164:90–100

    Article  PubMed  CAS  Google Scholar 

  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 183:56–60

    PubMed  CAS  Google Scholar 

  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 183:583–588

    PubMed  CAS  Google Scholar 

  11. Watanabe Y, Toki A, Todani T (1999) Bile duct cancer developed after cyst excision for choledochal cyst. J Hepatobil Pancreat Surg 6:207–212

    Article  CAS  Google Scholar 

  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 126:939–944

    Article  PubMed  CAS  Google Scholar 

  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 36:856–862

    Article  PubMed  CAS  Google Scholar 

  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 10:1183–1187

    PubMed  Google Scholar 

  15. Ando H, Ito T, Watanabe Y, Seo T, Kaneko K, Nagaya M (1995) Spontaneous perforation of choledochal cyst. J Am Coll Surg 181:125–128

    PubMed  CAS  Google Scholar 

  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 30:1658–1662

    Article  PubMed  CAS  Google Scholar 

  17. Shevchenko A, Wilm M, Vorm O, Mann M (1996) Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal Chem 68:850–858

    Article  PubMed  CAS  Google Scholar 

  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 84:100–106

    Article  PubMed  CAS  Google Scholar 

  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 265:7432–7439

    PubMed  CAS  Google Scholar 

  20. Patard L, Lallemand JY, Stoven V (2003) An insight into the role of human pancreatic lithostathine. JOP 4:92–103

    PubMed  Google Scholar 

  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 4:680–689

    Article  PubMed  CAS  Google Scholar 

  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 99:1421–1430

    PubMed  CAS  Google Scholar 

  23. Lee BI, Mustafi D, Cho W, Nakagawa Y (2003) Characterization of calcium binding properties of lithostathine. J Biol Inorg Chem 8:341–347

    PubMed  CAS  Google Scholar 

  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 20:3313–3321

    Article  PubMed  CAS  Google Scholar 

  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 276:21028–21038

    Article  PubMed  CAS  Google Scholar 

  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 1723:8–18

    PubMed  CAS  Google Scholar 

  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 84:102–107

    PubMed  CAS  Google Scholar 

  28. Multigner L, Daudon M, Montalto G, De Caro A, Etienne JP, Sarles H (1986) Radiolucent pancreatic stones. N Engl J Med 314:248

    PubMed  CAS  Google Scholar 

  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 36:1509–1516

    Article  CAS  PubMed  Google Scholar 

  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 86:419–423

    Article  PubMed  CAS  Google Scholar 

  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 51:51770–51778

    Article  CAS  Google Scholar 

  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 22:79–88

    Article  PubMed  CAS  Google Scholar 

  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 16:26–28

    Article  PubMed  CAS  Google Scholar 

  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 3:715–728

    Article  PubMed  CAS  Google Scholar 

  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;3569–3578

    Article  PubMed  CAS  Google Scholar 

  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 3:1042–1055

    Article  PubMed  CAS  Google Scholar 

  37. He XM, Carter DC (1992) Atomic structure and chemistry of human serum albumin. Nature 358:209–215

    Article  PubMed  CAS  Google Scholar 

  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 45:52–58

    Article  PubMed  CAS  Google Scholar 

  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 61:862–866

    Article  PubMed  Google Scholar 

  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 49:1953–1956

    Article  PubMed  Google Scholar 

  41. Bensimon G, Lacomblez L, Meininger V (1994) A controlled trial of riluzole in amyotrophic lateral sclerosis. N Engl J Med 330:585–591

    Article  PubMed  CAS  Google Scholar 

  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 99:16400–16406

    Article  PubMed  CAS  Google Scholar 

  43. Huntington Study Group (2003) Dosage effects of riluzole in Huntington's disease. A multicenter placebo-controlled study. Neurology 61:1551–1556

    Google Scholar 

  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 26:1922–1928

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work has been supported by a grant-in-aid (17591860) for Scientific Research from the Ministry of Education, Science, and Culture, Japan. We thank Kenji Yamamoto, Ikuko Sagawa, and Satoru Kuboi (APRO Life Science Institute, Naruto, Japan) for peptide mass fingerprinting analysis.

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  1. Department of Pediatric Surgery, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8560, Japan

    Kenitiro Kaneko, Hisami Ando, Takahiko Seo, Yasuyuki Ono, Takahisa Tainaka & Wataru Sumida

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  1. Kenitiro Kaneko
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  2. Hisami Ando
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  3. Takahiko Seo
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Correspondence to Kenitiro Kaneko.

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Kaneko, K., Ando, H., Seo, T. et al. Proteomic Analysis of Protein Plugs: Causative Agent of Symptoms in Patients with Choledochal Cyst. Dig Dis Sci 52, 1979–1986 (2007). https://doi.org/10.1007/s10620-006-9398-4

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  • DOI: https://doi.org/10.1007/s10620-006-9398-4

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