Isolation of an acidic protein from cholesterol gallstones, which inhibits the precipitation of calcium carbonate in vitro

S. Shimizu, B. Sabsay, A. Veis, J. D. Ostrow, Robert V Rege, L. G. Dawes

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Abstract

In seeking to identify nucleating/antinucleating proteins involved in the pathogenesis of cholesterol gallstones, a major acidic protein was isolated from each of 13 samples of cholesterol gallstones. After the stones were extracted with methyl t-butyl ether to remove cholesterol, and methanol to remove bile salts and other lipids, they were demineralized with EDTA. The extracts were desalted with Sephadex-G25, and the proteins separated by PAGE. A protein was isolated, of molecular weight below 10 kD, which included firmly-bound diazo-positive yellow pigments and contained 24% acidic, but only 7% basic amino acid residues. The presence of N-acetyl glucosamine suggested that this was a glycoprotein. This protein at concentrations as low as 2 μg/ml, but neither human serum albumin nor its complex with bilirubin, inhibited calcium carbonate precipitation from a supersaturated solution in vitro. This protein could be precipitated from 0.15 M NaCl solution by the addition of 0.5 M calcium chloride. Considering that cholesterol gallstones contain calcium and pigment at their centers, and that small acidic proteins are important regulators in other biomineralization systems, this protein seems likely to play a role in the pathogenesis of cholesterol gallstones.

Original languageEnglish (US)
Pages (from-to)1990-1996
Number of pages7
JournalJournal of Clinical Investigation
Volume84
Issue number6
StatePublished - 1989

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Calcium Carbonate
Gallstones
Cholesterol
Proteins
Basic Amino Acids
Calcium Chloride
In Vitro Techniques
Glucosamine
Bile Acids and Salts
Bilirubin
Serum Albumin
Edetic Acid
Methanol
Glycoproteins
Molecular Weight
Calcium
Lipids

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Isolation of an acidic protein from cholesterol gallstones, which inhibits the precipitation of calcium carbonate in vitro. / Shimizu, S.; Sabsay, B.; Veis, A.; Ostrow, J. D.; Rege, Robert V; Dawes, L. G.

In: Journal of Clinical Investigation, Vol. 84, No. 6, 1989, p. 1990-1996.

Research output: Contribution to journalArticle

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AU - Rege, Robert V

AU - Dawes, L. G.

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AB - In seeking to identify nucleating/antinucleating proteins involved in the pathogenesis of cholesterol gallstones, a major acidic protein was isolated from each of 13 samples of cholesterol gallstones. After the stones were extracted with methyl t-butyl ether to remove cholesterol, and methanol to remove bile salts and other lipids, they were demineralized with EDTA. The extracts were desalted with Sephadex-G25, and the proteins separated by PAGE. A protein was isolated, of molecular weight below 10 kD, which included firmly-bound diazo-positive yellow pigments and contained 24% acidic, but only 7% basic amino acid residues. The presence of N-acetyl glucosamine suggested that this was a glycoprotein. This protein at concentrations as low as 2 μg/ml, but neither human serum albumin nor its complex with bilirubin, inhibited calcium carbonate precipitation from a supersaturated solution in vitro. This protein could be precipitated from 0.15 M NaCl solution by the addition of 0.5 M calcium chloride. Considering that cholesterol gallstones contain calcium and pigment at their centers, and that small acidic proteins are important regulators in other biomineralization systems, this protein seems likely to play a role in the pathogenesis of cholesterol gallstones.

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