Isolation of a human biliary glycoprotein inhibitor of cholesterol crystallization.
BACKGROUND
About 50% of populations in developed countries have bile supersaturated with cholesterol, which is a major risk factor for cholesterol gallstone formation. Despite the prevalence of supersaturated bile, only about 10% of these populations develop gallstones. The existence of a biliary protein that inhibits cholesterol crystallization was hypothesized to explain this discrepancy. This report outlines the purification and characterization of such a human biliary glycoprotein.


METHODS
Chromatographic methods were used for separation and characterization. Additional steps included activity analysis by crystal growth assay, electrophoresis, and deglycosylation.


RESULTS
The glycoprotein consists of a heterodimer, M(r) of 120 kilodalton, with subunits of M(r) of 63 kilodalton and 58 kilodalton. Each of the subunits is characterized by an isoelectric point of 6.6 and shows comparable inhibitory activity. Deglycosylation of the subunits show that they share a similar polypeptide backbone (M(r) of 35 kilodalton) based upon a highly similar amino acid profile. This suggests that differential subunit glycosylation alone may account for the apparent heterodimeric structure.


CONCLUSIONS
No other human biliary glycoprotein has been found thus far that shows cholesterol crystal growth-inhibiting activity. Thus, it may be of importance in preventing gallstone formation in healthy populations.
Ohya
T
T
Gastrointestinal Research Unit, Cleveland Clinic Foundation, Ohio.
Schwarzendrube
J
J
Busch
N
N
Gresky
S
S
Chandler
K
K
Takabayashi
A
A
Igimi
H
H
Egami
K
K
Holzbach
R T
RT
D000596:Amino Acids / Q000032:analysis; D001646:Bile / Q000737:chemistry*; D002769:Cholelithiasis / Q000209:etiology; D002784:Cholesterol / Q000378:metabolism*; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003460:Crystallization; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000032:analysis / Q000302:isolation & purification* / Q000494:pharmacology; D006031:Glycosylation; D006801:Humans
D000596:Amino Acids; D006023:Glycoproteins; D002784:Cholesterol