Human endothelial cells synthesize, process, and secrete fibronectin molecules bearing an alternatively spliced type III homology (ED1)

John H. Peters, Lee Ann Sporn, Mark H. Ginsberg, Denisa D. Wagner

Research output: Contribution to journalArticle

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Abstract

Cellular fibronectin (Fn) bearing an alternatively spliced extra type III structural repeat (ED1) is normally present at low concentrations in blood plasma. The source of this material remains uncertain. In this study, primary cultures of human umbilical vein endothelial cells (HUVEC) labeled with 35S-methionine were observed to synthesize Fn monomers both with and without this segment. Monomers isolated from cell lysates with antibodies to the ED1 sequence comigrated in nonreduced sodium dodecyl sulfate polyacrylamide gel electrophoresis with the slower (designated M1 ), but not the faster (designated M2), of two major monomeric populations that were recognized by antibodies raised to plasma-derived Fn. The differences between M1 and M2 were not due to glycosylation, since they were also observed between species of Fn monomer purified from cells grown in the presence of tunicamycin. M1 and M2 were both observed to incorporate with a similar rate into dimeric Fn, indicating that Fn monomers with and without the ED1 site can dimerize with similar efficiency. Analysis of reduced samples of Fn isolated from cells with anti-ED1 antibodies indicated the presence of both M1-M1 and M1-M2 dimers. In addition to being incorporated into extracellular matrix, ED1 + Fn was observed to be secreted in soluble form into the medium, potentially reflecting intravascular release of this protein by endothelial cells in vivo.

Original languageEnglish (US)
Pages (from-to)1801-1808
Number of pages8
JournalBlood
Volume75
Issue number9
StatePublished - May 1 1990
Externally publishedYes

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Bearings (structural)
Endothelial cells
Fibronectins
Endothelial Cells
Molecules
Monomers
Antibodies
Glycosylation
Plasmas
Tunicamycin
Human Umbilical Vein Endothelial Cells
Electrophoresis
Cell culture
Sodium Dodecyl Sulfate
Methionine
Dimers
Extracellular Matrix
Polyacrylamide Gel Electrophoresis
Anti-Idiotypic Antibodies
Blood

ASJC Scopus subject areas

  • Hematology

Cite this

Human endothelial cells synthesize, process, and secrete fibronectin molecules bearing an alternatively spliced type III homology (ED1). / Peters, John H.; Sporn, Lee Ann; Ginsberg, Mark H.; Wagner, Denisa D.

In: Blood, Vol. 75, No. 9, 01.05.1990, p. 1801-1808.

Research output: Contribution to journalArticle

Peters, John H. ; Sporn, Lee Ann ; Ginsberg, Mark H. ; Wagner, Denisa D. / Human endothelial cells synthesize, process, and secrete fibronectin molecules bearing an alternatively spliced type III homology (ED1). In: Blood. 1990 ; Vol. 75, No. 9. pp. 1801-1808.
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