Cholesterol crystallization‐promoting activity of aminopeptidase‐N isolated from the vesicular carrier of biliary lipids

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Abstract
Different hydrophobic glycoproteins are associated to native biliary vesicles, which are the major carrier of biliary cholesterol. Some of these proteins promote cholesterol crystallization, a key step in cholesterol gallstone formation. This study was specifically conducted to identify the 130 kDa biliary vesicle‐associated glycoprotein and to determine its in vitro effect on the cholesterol crystal formation time. The 130 kDa vesicular glycoprotein was identified as aminopeptidase‐N by amino acid sequencing and specific enzymatic assay. Polyclonal antibodies raised against aminopeptidase‐N allowed us to determine its concentration in human hepatic bile, which varied from 17.3 to 57.6 μg/ml. Aminopeptidase‐N showed a concentration‐dependent cholesterol crystallization activity when it was added to supersaturated model bile at a concentration range usually found in native bile. Because of this promoting effect on in vitro cholesterol crystal formation, we suggest that biliary aminopeptidase‐N may play a critical role in the pathogenesis of cholesterol gallstone disease.