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

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.

KW - Animals

KW - Cattle

KW - Cells, Cultured

KW - Chromatography, Affinity

KW - Electrophoresis, Polyacrylamide Gel

KW - Endothelium/metabolism

KW - Eye Proteins/biosynthesis

KW - Isoelectric Focusing

KW - Molecular Weight

KW - Peptide Mapping

KW - Retina/metabolism

U2 - 10.1042/bj2460121

DO - 10.1042/bj2460121

M3 - Article

C2 - 3675551

SN - 0264-6021

VL - 246

SP - 121

EP - 129

JO - The Biochemical journal

JF - The Biochemical journal

IS - 1

ER -