Skip to main content
PMC home page
Gut logoLink to Gut
. 1995 Sep;37(3):422–426. doi: 10.1136/gut.37.3.422

Biliary aminopeptidase-N and the cholesterol crystallisation defect in cholelithiasis.

L Núñez 1, L Amigo 1, G Mingrone 1, A Rigotti 1, L Puglielli 1, A Raddatz 1, F Pimentel 1, A V Greco 1, S González 1, J Garrido 1, et al.
PMCID: PMC1382827  PMID: 7590442

Abstract

Several biliary proteins have cholesterol crystallisation promoting activity. One of these glycoproteins is aminopeptidase-N, a canalicular ectoenzyme. This study attempted to localise aminopeptidase-N along the biliary tree, to assess its concentration in a series of 98 patients subjected to abdominal surgery, 40 of them without gall stones, and to correlate its concentration with cholesterol crystal formation time of gall bladder bile. Aminopeptidase-N was isolated from purified native biliary vesicles. A specific polyclonal rabbit anti-aminopeptidase-N antibody was prepared for quantitative immunoblotting and for immunolocalisation. Tissue was obtained from liver biopsy specimens and from gall bladders removed at surgery because of gall stone disease. Aminopeptidase-N was immunolocalised to the apical membranes of hepatocytes and to the apical pole of ductular and gall bladder mucosal cells. The nucleation time of gall bladder bile was mean (SD) 4 (3) days in the gall stone group, compared with 21 (18) days in the control group (p < 0.001). Total absolute biliary protein and aminopeptidase-N concentrations were similar in both the control and gall stone patients. There was a reciprocal significant correlation, however, between the nucleation time and the relative aminopeptidase-N concentration (r = -0.35, p < 0.01) only in the gall stone group of patients. This study shows that this apical transmembrane ectoenzyme with cholesterol crystallisation promoting activity is present along the biliary tree and the hepatocyte. These findings support the concept that high concentrations or qualitative changes of biliary aminopeptidase-N contribute to cholesterol gall stone formation.

Full text

PDF
422

Images in this article

423Figure 1
on p.423
424Figure 2
on p.424

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abei M., Kawczak P., Nuutinen H., Langnas A., Svanvik J., Holzbach R. T. Isolation and characterization of a cholesterol crystallization promoter from human bile. Gastroenterology. 1993 Feb;104(2):539–548. doi: 10.1016/0016-5085(93)90424-b. [DOI] [PubMed] [Google Scholar]
  2. Abei M., Schwarzendrube J., Nuutinen H., Broughan T. A., Kawczak P., Williams C., Holzbach R. T. Cholesterol crystallization-promoters in human bile: comparative potencies of immunoglobulins, alpha 1-acid glycoprotein, phospholipase C, and aminopeptidase N1. J Lipid Res. 1993 Jul;34(7):1141–1148. [PubMed] [Google Scholar]
  3. Burnstein M. J., Ilson R. G., Petrunka C. N., Taylor R. D., Strasberg S. M. Evidence for a potent nucleating factor in the gallbladder bile of patients with cholesterol gallstones. Gastroenterology. 1983 Oct;85(4):801–807. [PubMed] [Google Scholar]
  4. Chijiiwa K., Koga A., Yamasaki T., Shimada K., Noshiro H., Nakayama F. Fibronectin: a possible factor promoting cholesterol monohydrate crystallization in bile. Biochim Biophys Acta. 1991 Oct 15;1086(1):44–48. doi: 10.1016/0005-2760(91)90152-8. [DOI] [PubMed] [Google Scholar]
  5. Gallinger S., Harvey P. R., Petrunka C. N., Ilson R. G., Strasberg S. M. Biliary proteins and the nucleation defect in cholesterol cholelithiasis. Gastroenterology. 1987 Apr;92(4):867–875. doi: 10.1016/0016-5085(87)90959-0. [DOI] [PubMed] [Google Scholar]
  6. Gollish S. H., Burnstein M. J., Ilson R. G., Petrunka C. N., Strasberg S. M. Nucleation of cholesterol monohydrate crystals from hepatic and gall-bladder bile of patients with cholesterol gall stones. Gut. 1983 Sep;24(9):836–844. doi: 10.1136/gut.24.9.836. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Groen A. K., Noordam C., Drapers J. A., Egbers P., Jansen P. L., Tytgat G. N. Isolation of a potent cholesterol nucleation-promoting activity from human gallbladder bile: role in the pathogenesis of gallstone disease. Hepatology. 1990 Apr;11(4):525–533. doi: 10.1002/hep.1840110402. [DOI] [PubMed] [Google Scholar]
  8. Groen A. K., Stout J. P., Drapers J. A., Hoek F. J., Grijm R., Tytgat G. N. Cholesterol nucleation-influencing activity in T-tube bile. Hepatology. 1988 Mar-Apr;8(2):347–352. doi: 10.1002/hep.1840080226. [DOI] [PubMed] [Google Scholar]
  9. Harvey P. R., Strasberg S. M. Will the real cholesterol-nucleating and -antinucleating proteins please stand up? Gastroenterology. 1993 Feb;104(2):646–650. doi: 10.1016/0016-5085(93)90439-j. [DOI] [PubMed] [Google Scholar]
  10. Harvey P. R., Upadhya G. A., Strasberg S. M. Immunoglobulins as nucleating proteins in the gallbladder bile of patients with cholesterol gallstones. J Biol Chem. 1991 Jul 25;266(21):13996–14003. [PubMed] [Google Scholar]
  11. Holan K. R., Holzbach R. T., Hermann R. E., Cooperman A. M., Claffey W. J. Nucleation time: a key factor in the pathogenesis of cholesterol gallstone disease. Gastroenterology. 1979 Oct;77(4 Pt 1):611–617. [PubMed] [Google Scholar]
  12. Inokuchi K. Improved survival after prophylactic portal nondecompression surgery for esophageal varices: a randomized clinical trial. Cooperative Study Group of Portal Hypertension of Japan. Hepatology. 1990 Jul;12(1):1–6. doi: 10.1002/hep.1840120102. [DOI] [PubMed] [Google Scholar]
  13. Kibe A., Holzbach R. T., LaRusso N. F., Mao S. J. Inhibition of cholesterol crystal formation by apolipoproteins in supersaturated model bile. Science. 1984 Aug 3;225(4661):514–516. doi: 10.1126/science.6429856. [DOI] [PubMed] [Google Scholar]
  14. Levy P. F., Smith B. F., LaMont J. T. Human gallbladder mucin accelerates nucleation of cholesterol in artificial bile. Gastroenterology. 1984 Aug;87(2):270–275. [PubMed] [Google Scholar]
  15. Markwell M. A., Haas S. M., Bieber L. L., Tolbert N. E. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem. 1978 Jun 15;87(1):206–210. doi: 10.1016/0003-2697(78)90586-9. [DOI] [PubMed] [Google Scholar]
  16. Miquel J. F., Núez L., Rigotti A., Amigo L., Brandan E., Nervi F. Isolation and partial characterization of cholesterol pronucleating hydrophobic glycoproteins associated to native biliary vesicles. FEBS Lett. 1993 Feb 22;318(1):45–49. doi: 10.1016/0014-5793(93)81324-s. [DOI] [PubMed] [Google Scholar]
  17. Miquel J. F., Rigotti A., Rojas E., Brandan E., Nervi F. Isolation and purification of human biliary vesicles with potent cholesterol-nucleation-promoting activity. Clin Sci (Lond) 1992 Feb;82(2):175–180. doi: 10.1042/cs0820175. [DOI] [PubMed] [Google Scholar]
  18. Offner G. D., Gong D., Afdhal N. H. Identification of a 130-kilodalton human biliary concanavalin A binding protein as aminopeptidase N. Gastroenterology. 1994 Mar;106(3):755–762. doi: 10.1016/0016-5085(94)90712-9. [DOI] [PubMed] [Google Scholar]
  19. Ohya T., Schwarzendrube J., Busch N., Gresky S., Chandler K., Takabayashi A., Igimi H., Egami K., Holzbach R. T. Isolation of a human biliary glycoprotein inhibitor of cholesterol crystallization. Gastroenterology. 1993 Feb;104(2):527–538. doi: 10.1016/0016-5085(93)90423-a. [DOI] [PubMed] [Google Scholar]
  20. Rigotti A., Núez L., Amigo L., Puglielli L., Garrido J., Santos M., González S., Mingrone G., Greco A., Nervi F. Biliary lipid secretion: immunolocalization and identification of a protein associated with lamellar cholesterol carriers in supersaturated rat and human bile. J Lipid Res. 1993 Nov;34(11):1883–1894. [PubMed] [Google Scholar]
  21. Roman L. M., Hubbard A. L. A domain-specific marker for the hepatocyte plasma membrane: localization of leucine aminopeptidase to the bile canalicular domain. J Cell Biol. 1983 Jun;96(6):1548–1558. doi: 10.1083/jcb.96.6.1548. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Smith B. F., LaMont J. T. Hydrophobic binding properties of bovine gallbladder mucin. J Biol Chem. 1984 Oct 10;259(19):12170–12177. [PubMed] [Google Scholar]
  23. Upadhya G. A., Harvey P. R., Strasberg S. M. Effect of human biliary immunoglobulins on the nucleation of cholesterol. J Biol Chem. 1993 Mar 5;268(7):5193–5200. [PubMed] [Google Scholar]

Articles from Gut are provided here courtesy of BMJ Publishing Group

RESOURCES