Isolation and characterization of a 115,000-dalton matrix-associated glycoprotein from chick aorta

G. M. Bressan, I. Castellani, A. Colombatti, D. Volpin

Research output: Contribution to journalArticlepeer-review

Abstract

Chick aortas were extracted sequentially phosphate-buffered saline, 6 M guanidine HCl, and 6 M guanidine HCl containing dithioerythritol. The proteins present in the guanidine HCl + dithioerythritrol extract were separated by DEAE-cellulose chromatography and the fractions recovered were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Five major glycoprotein components with apparent M(r) = 205,000, 195,000, 150,000, 135,000, and 115,000 (gp 115) were identified. gp 115 was further studied since it was the only noncollagenous protein based on amino acid analysis. The protein was purified to homogeneity by preparative electrophoresis. Its amino acid composition was characterized by a high content of glutamic acid and arginine and a relatively high content of leucine, glycine and alanine. The concentration of gp 115 in the guanidine HCl + dithioerythritol extract was about 15-fold that in the guanidine and saline extracts. Overall, about 80% of the protein was solubilized with guanidine HCl + dithioerythritol, suggesting that most of it formed large aggregates stabilized by disulfide bonds in vivo. Immunofluorescence studies with specific antibodies showed that gp 115 formed an extracellular fibrillar network in the aorta wall. One-dimensional finger printing with Staphylococcus aureus V8 protease and immunological studies indicated that the protein was unrelated to fibronectin and laminin. The data led us to conclude that gp 115 is a novel extracellular component of chick aorta.

Original languageEnglish
Pages (from-to)13262-13267
Number of pages6
JournalJournal of Biological Chemistry
Volume258
Issue number21
Publication statusPublished - 1983

ASJC Scopus subject areas

  • Biochemistry

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