Identification and partial characterization of two major proteins of Mr 47,000 synthesized by bovine retinal endothelial cells in culture

A E Canfield, A M Schor, D C West, S L Schor, M E Grant

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Biosynthetic experiments with cultured bovine retinal endothelial cells have identified a glycoprotein of Mr 47,000 (Gp47) as a major component secreted into the medium. Gp47 is a non-collagenous glycoprotein with a pI of 4.6-5.5, which does not bind to either gelatin-Sepharose or heparin-Sepharose but is retained by concanavalin A-Sepharose. The Mr of this species decreases to approx. 42,000 in the presence of tunicamycin, indicating that it contains asparagine-linked oligosaccharides. A second protein of Mr 47,000 (P47) is present in the cell layer/matrix of these cultured cells. The electrophoretic mobility of P47 remains unaltered when synthesized in the presence of tunicamycin. Peptide-mapping experiments using N-chlorosuccinimide and Staphylococcus aureus V8 proteinase demonstrate that Gp47 and P47 are distinct proteins, and are not related to colligin, a membrane-bound collagen-receptor protein of similar size, or to SPARC, a major secreted product of parietal endodermal cells and sparse cultures of aortic endothelial cells.

Original languageEnglish
Pages (from-to)121-9
Number of pages9
JournalThe Biochemical journal
Volume246
Issue number1
DOIs
Publication statusPublished - 15 Aug 1987

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Endothelial cells
Cell culture
Glycoproteins
Endothelial Cells
Cell Culture Techniques
Tunicamycin
Proteins
Cells
Collagen Receptors
Electrophoretic mobility
Peptide Mapping
Asparagine
Gelatin
Oligosaccharides
Sepharose
Staphylococcus aureus
Cultured Cells
Peptide Hydrolases
Experiments
Membranes

Keywords

  • Animals
  • Cattle
  • Cells, Cultured
  • Chromatography, Affinity
  • Electrophoresis, Polyacrylamide Gel
  • Endothelium/metabolism
  • Eye Proteins/biosynthesis
  • Isoelectric Focusing
  • Molecular Weight
  • Peptide Mapping
  • Retina/metabolism

Cite this

Canfield, A E ; Schor, A M ; West, D C ; Schor, S L ; Grant, M E. / Identification and partial characterization of two major proteins of Mr 47,000 synthesized by bovine retinal endothelial cells in culture. In: The Biochemical journal. 1987 ; Vol. 246, No. 1. pp. 121-9.
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abstract = "Biosynthetic experiments with cultured bovine retinal endothelial cells have identified a glycoprotein of Mr 47,000 (Gp47) as a major component secreted into the medium. Gp47 is a non-collagenous glycoprotein with a pI of 4.6-5.5, which does not bind to either gelatin-Sepharose or heparin-Sepharose but is retained by concanavalin A-Sepharose. The Mr of this species decreases to approx. 42,000 in the presence of tunicamycin, indicating that it contains asparagine-linked oligosaccharides. A second protein of Mr 47,000 (P47) is present in the cell layer/matrix of these cultured cells. The electrophoretic mobility of P47 remains unaltered when synthesized in the presence of tunicamycin. Peptide-mapping experiments using N-chlorosuccinimide and Staphylococcus aureus V8 proteinase demonstrate that Gp47 and P47 are distinct proteins, and are not related to colligin, a membrane-bound collagen-receptor protein of similar size, or to SPARC, a major secreted product of parietal endodermal cells and sparse cultures of aortic endothelial cells.",
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Identification and partial characterization of two major proteins of Mr 47,000 synthesized by bovine retinal endothelial cells in culture. / Canfield, A E; Schor, A M; West, D C; Schor, S L; Grant, M E.

In: The Biochemical journal, Vol. 246, No. 1, 15.08.1987, p. 121-9.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Identification and partial characterization of two major proteins of Mr 47,000 synthesized by bovine retinal endothelial cells in culture

AU - Canfield, A E

AU - Schor, A M

AU - West, D C

AU - Schor, S L

AU - Grant, M E

PY - 1987/8/15

Y1 - 1987/8/15

N2 - Biosynthetic experiments with cultured bovine retinal endothelial cells have identified a glycoprotein of Mr 47,000 (Gp47) as a major component secreted into the medium. Gp47 is a non-collagenous glycoprotein with a pI of 4.6-5.5, which does not bind to either gelatin-Sepharose or heparin-Sepharose but is retained by concanavalin A-Sepharose. The Mr of this species decreases to approx. 42,000 in the presence of tunicamycin, indicating that it contains asparagine-linked oligosaccharides. A second protein of Mr 47,000 (P47) is present in the cell layer/matrix of these cultured cells. The electrophoretic mobility of P47 remains unaltered when synthesized in the presence of tunicamycin. Peptide-mapping experiments using N-chlorosuccinimide and Staphylococcus aureus V8 proteinase demonstrate that Gp47 and P47 are distinct proteins, and are not related to colligin, a membrane-bound collagen-receptor protein of similar size, or to SPARC, a major secreted product of parietal endodermal cells and sparse cultures of aortic endothelial cells.

AB - Biosynthetic experiments with cultured bovine retinal endothelial cells have identified a glycoprotein of Mr 47,000 (Gp47) as a major component secreted into the medium. Gp47 is a non-collagenous glycoprotein with a pI of 4.6-5.5, which does not bind to either gelatin-Sepharose or heparin-Sepharose but is retained by concanavalin A-Sepharose. The Mr of this species decreases to approx. 42,000 in the presence of tunicamycin, indicating that it contains asparagine-linked oligosaccharides. A second protein of Mr 47,000 (P47) is present in the cell layer/matrix of these cultured cells. The electrophoretic mobility of P47 remains unaltered when synthesized in the presence of tunicamycin. Peptide-mapping experiments using N-chlorosuccinimide and Staphylococcus aureus V8 proteinase demonstrate that Gp47 and P47 are distinct proteins, and are not related to colligin, a membrane-bound collagen-receptor protein of similar size, or to SPARC, a major secreted product of parietal endodermal cells and sparse cultures of aortic endothelial cells.

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KW - Isoelectric Focusing

KW - Molecular Weight

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KW - Retina/metabolism

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