Inhibition and promotion of cholesterol crystallization by protein fractions from normal human gallbladder bile.
Pooled, normal human gallbladder biles were initially separated on a molecular sieving chromatography column to remove soluble mucin glycoproteins as well as high molecular weight proteins (greater than 200,000). The remaining lower molecular weight proteins and other bile components were then examined by lectin affinity chromatography with four different types of lectin. The separated bound fractions were compared for inhibiting and promoting activities with a newly devised sensitive cholesterol crystal growth assay and for differences in electrophoretic patterns on SDS-gels. Protein factors (presumably glycoproteins) were found to have both inhibiting and promoting activities, even in the absence of cholesterol gallstone disease. The promoting effect was indicated by shortened crystal detection times and increases in crystal growth rate; whereas the inhibiting effect was indicated by decreases in crystal growth rate and reductions in the final crystal concentration as determined by the growth assay. Affinity chromatography mitigated the major problems of removing both lipids and pigment from the glycoproteins. In addition, partial purification of bound fractions with potent cholesterol crystal nucleation-altering activity can be obtained by this technique.
Busch
N
N
Department of Gastroenterology, Cleveland Clinic Foundation, OH 44195-5218.
Matiuck
N
N
Sahlin
S
S
Pillay
S P
SP
Holzbach
R T
RT
D000924:Anticholesteremic Agents / Q000494:pharmacology*; D001646:Bile / Q000737:chemistry / Q000502:physiology*; D002784:Cholesterol / Q000737:chemistry / Q000378:metabolism*; D002846:Chromatography, Affinity; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D005704:Gallbladder / Q000737:chemistry / Q000502:physiology*; D006023:Glycoproteins / Q000502:physiology*; D006801:Humans; D008970:Molecular Weight; D012835:Silver Nitrate; D014961:X-Ray Diffraction
D000924:Anticholesteremic Agents; D006023:Glycoproteins; D012835:Silver Nitrate; D002784:Cholesterol