Identification of human biliary alpha 1-acid glycoprotein as a cholesterol crystallization promoter.
OBJECTIVE
We have recently outlined the biochemical features of a human 42-kilodalton biliary glycoprotein that shows concentration-dependent cholesterol crystallization-promoting activity. The goal in this work was to establish its identity and to examine some aspects of its biochemical properties relative to its activity.


METHODS
Internal amino acid sequencing following tryptic digestion was performed. Based upon this result, immunoreactivity against the 42-kilodalton glycoprotein was examined using a relevant antibody. With the same antibody, the 42-kilodalton glycoprotein was isolated from bile and assayed for activity. Sequential enzymatic deglycosylation of successive terminal glycans of the purified glycoprotein was performed, and the effects on both reductions in molecular radius (M(r)) and on comparative promoter activities were examined.


RESULTS
Both amino acid sequence and immunochemical data identify the 42-kilodalton glycoprotein as a biliary form of alpha 1-acid glycoprotein. When purified by immunoaffinity chromatography, potent promoting activity shown was proportionately reduced by successive removal of terminal glycans that also reduced the M(r)s.


CONCLUSIONS
The 42-kilodalton cholesterol crystallization-promoting glycoprotein is now identified as a biliary form of alpha 1-acid glycoprotein. Further, some aspects of the important role of glycans in this extensively glycosylated protein have been explored.
Abei
M
M
Gastrointestinal Research Unit, Cleveland Clinic Foundation, Ohio.
Nuutinen
H
H
Kawczak
P
P
Schwarzendrube
J
J
Pillay
S P
SP
Holzbach
R T
RT
D001646:Bile / Q000737:chemistry* / Q000187:drug effects; D002784:Cholesterol / Q000737:chemistry / Q000502:physiology*; D003208:Concanavalin A / Q000494:pharmacology; D003460:Crystallization; D006801:Humans; D007120:Immunochemistry; D008970:Molecular Weight; D009961:Orosomucoid / Q000378:metabolism / Q000494:pharmacology*
D009961:Orosomucoid; D003208:Concanavalin A; D002784:Cholesterol