Immunoglobulins as nucleating proteins in the gallbladder bile of patients with cholesterol gallstones
| Autor: | Steven M. Strasberg, P. R. C. Harvey, G A Upadhya |
|---|---|
| Rok vydání: | 1991 |
| Předmět: |
Gel electrophoresis
chemistry.chemical_classification biology Cholesterol Gallbladder Fast protein liquid chromatography Cell Biology Biochemistry chemistry.chemical_compound medicine.anatomical_structure chemistry Affinity chromatography Concanavalin A Electroelution biology.protein medicine Glycoprotein Molecular Biology |
| Zdroj: | Journal of Biological Chemistry. 266:13996-14003 |
| ISSN: | 0021-9258 |
| Popis: | The gallbladder bile of patients with cholesterol gallstones contains pronucleating proteins which accelerate precipitation of cholesterol crystals from bile. In this study we have improved the purification procedure developed earlier for these nucleating proteins and have now identified the nature of these proteins. Gallbladder bile from patients with cholesterol gallstones was applied to concanavalin A affinity columns. The ConA-binding glycoprotein fractions containing the nucleating proteins were then separated by FPLC (fast protein liquid chromatography) using a Superose 12 gel filtration column. Nucleating activity was detected in the high molecular weight (FPLC-1) as well as in the low molecular weight fractions (FPLC-3). Investigation of the high molecular weight fraction by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by electroelution and amino acid sequencing suggested that these proteins were immunoglobulins. Immunostaining of Western blots with specific monoclonal antibodies identified the presence of immunoglobulin (Ig) M and IgA in the FPLC-1 fraction. These immunoglobulins were further purified by affinity chromatography employing an antibody exchanger (ABx) column which specifically binds immunoglobulins. There was no reduction in the cholesterol nucleating activity in the Abx-bound fraction compared to FPLC-1. Additional studies showed that the FPLC-1 fraction was significantly more potent than the ConA glycoproteins from either rapid and slow nucleating biles. Also the number of crystals formed was significantly greater in the FPLC-1 fraction isolated from cholesterol gallstone biles than from the FPLC-1 fraction from control patient biles. Commercially obtained IgM and IgA had no effect on nucleation, but IgM isolated from the serum of patients with Waldenstrom's macroglobulinemia did accelerate the nucleation of cholesterol. We conclude that the IgM and possibly IgA are pronucleating proteins and may be important in the pathogenesis of cholesterol gallstones in man. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|