Isolation and purification of collagen α1(I) receptor from human platelet membrane

T. M. Chiang, Andrew Kang

Research output: Contribution to journalArticle

80 Citations (Scopus)

Abstract

We have previously demonstrated that chick-skin type I collagen and the α1(I) chain mediate platelet aggregation. Aggregation was associated with specific binding of these substances by platelet membranes. We now describe the isolation and purification of the receptor from isolated human platelet membrane. The receptor can be solubilized from platelet membranes with 0.5% Triton or 0.5% sodium deoxycholate. Using the combination of gel filtration, affinity column chromatography on α1-Sepharose 4B, and preparative slab gel electrophoresis, the receptor protein can be purified to a single band as judged on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Its activity was destroyed by incubation with trypsin or Pronase. The apparent molecular weight was estimated to be 65,000 by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Bound {14C]glycine-labeled α1(I) is displaced by unlabeled α1 chain. The binding of [14C] glycine-labeled α1(I) by the purified α1(I) receptor can also be inhibited by the receptor isolated from type I fibrillar collagen-Sepharose affinity chromatography. The data suggest that the α1(I) binding site is identical with the type I fibrillar collagen binding site.

Original languageEnglish (US)
Pages (from-to)7581-7586
Number of pages6
JournalJournal of Biological Chemistry
Volume257
Issue number13
StatePublished - Dec 1 1982

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Platelets
Fibrillar Collagens
Purification
Collagen
Blood Platelets
Electrophoresis
Membranes
Affinity Chromatography
Affinity chromatography
Sodium Dodecyl Sulfate
Glycine
Polyacrylamide Gel Electrophoresis
Binding Sites
Sepharose
Agarose Chromatography
Pronase
Deoxycholic Acid
Agglomeration
Gels
Collagen Type I

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Isolation and purification of collagen α1(I) receptor from human platelet membrane. / Chiang, T. M.; Kang, Andrew.

In: Journal of Biological Chemistry, Vol. 257, No. 13, 01.12.1982, p. 7581-7586.

Research output: Contribution to journalArticle

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AB - We have previously demonstrated that chick-skin type I collagen and the α1(I) chain mediate platelet aggregation. Aggregation was associated with specific binding of these substances by platelet membranes. We now describe the isolation and purification of the receptor from isolated human platelet membrane. The receptor can be solubilized from platelet membranes with 0.5% Triton or 0.5% sodium deoxycholate. Using the combination of gel filtration, affinity column chromatography on α1-Sepharose 4B, and preparative slab gel electrophoresis, the receptor protein can be purified to a single band as judged on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Its activity was destroyed by incubation with trypsin or Pronase. The apparent molecular weight was estimated to be 65,000 by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Bound {14C]glycine-labeled α1(I) is displaced by unlabeled α1 chain. The binding of [14C] glycine-labeled α1(I) by the purified α1(I) receptor can also be inhibited by the receptor isolated from type I fibrillar collagen-Sepharose affinity chromatography. The data suggest that the α1(I) binding site is identical with the type I fibrillar collagen binding site.

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