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Characterization of two monoclonal antibodies reactive with the external domain of the platelet-derived growth factor receptor

Rönnstrand, Lars LU ; Terracio, Louis; Claesson-Welsh, Lena; Heldin, Carl-Henrik and Rubin, Kristofer (1988) In Journal of Biological Chemistry 263(21). p.10429-10435
Abstract
Two monoclonal antibodies against the receptor for platelet-derived growth factor (PDGF) were obtained by immunizing mice with pure PDGF receptor preparations derived from porcine uterus. The antibodies, denoted PDGFR-B1 and PDGFR-B2, both bound to the external domain of the receptor, as demonstrated by indirect immunofluorescence and binding of 125I-labeled antibodies to intact human fibroblasts. Both antibodies precipitated pure 175-kDa 32P-labeled autophosphorylated porcine PDGF receptor as well as a Mr 175,000 glycoprotein from metabolically labeled cells. The monoclonal antibodies did not inhibit binding of 125I-PDGF to human fibroblasts and did not stimulate these cells to undergo mitosis. Both antibodies induced clustering and... (More)
Two monoclonal antibodies against the receptor for platelet-derived growth factor (PDGF) were obtained by immunizing mice with pure PDGF receptor preparations derived from porcine uterus. The antibodies, denoted PDGFR-B1 and PDGFR-B2, both bound to the external domain of the receptor, as demonstrated by indirect immunofluorescence and binding of 125I-labeled antibodies to intact human fibroblasts. Both antibodies precipitated pure 175-kDa 32P-labeled autophosphorylated porcine PDGF receptor as well as a Mr 175,000 glycoprotein from metabolically labeled cells. The monoclonal antibodies did not inhibit binding of 125I-PDGF to human fibroblasts and did not stimulate these cells to undergo mitosis. Both antibodies induced clustering and down-regulation of their antigen. However, this resulted in only a partial loss of cell surface binding sites for PDGF itself, consistent with the conclusion that the monoclonals recognized only one of two or several receptors for PDGF. Clustering and down-regulation were not seen when the cells were incubated with monovalent Fab' fragments of the PDGFR-B2 antibody. The antibodies also stimulated autophosphorylation of pure PDGF receptor, and PDGFR-B2 was shown to stimulate phosphorylation of phosphofructokinase, an exogenous substrate for the PDGF receptor kinase. High concentrations of PDGFR-B2 antibody, or Fab' fragments thereof, failed to enhance the PDGF receptor kinase activity, compatible with the possibility that dimerization was of importance in the antibody-stimulated kinase activity of purified PDGF receptors. (Less)
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author
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Animals *Antibodies, Cell Surface/*immunology/metabolism Receptors, Monoclonal Antigen-Antibody Complex Enzyme-Linked Immunosorbent Assay Epitopes/*analysis Female Kinetics Platelet-Derived Growth Factor/metabolism Receptors, Platelet-Derived Growth Factor Swine Uterus/metabolism
in
Journal of Biological Chemistry
volume
263
issue
21
pages
10429 - 10435
publisher
ASBMB
external identifiers
  • scopus:0023694016
ISSN
1083-351X
language
English
LU publication?
no
id
64336f64-e26e-4b85-ac1e-2e8a619bc66f (old id 1784230)
date added to LUP
2011-02-07 15:17:13
date last changed
2017-08-06 04:44:38
@article{64336f64-e26e-4b85-ac1e-2e8a619bc66f,
  abstract     = {Two monoclonal antibodies against the receptor for platelet-derived growth factor (PDGF) were obtained by immunizing mice with pure PDGF receptor preparations derived from porcine uterus. The antibodies, denoted PDGFR-B1 and PDGFR-B2, both bound to the external domain of the receptor, as demonstrated by indirect immunofluorescence and binding of 125I-labeled antibodies to intact human fibroblasts. Both antibodies precipitated pure 175-kDa 32P-labeled autophosphorylated porcine PDGF receptor as well as a Mr 175,000 glycoprotein from metabolically labeled cells. The monoclonal antibodies did not inhibit binding of 125I-PDGF to human fibroblasts and did not stimulate these cells to undergo mitosis. Both antibodies induced clustering and down-regulation of their antigen. However, this resulted in only a partial loss of cell surface binding sites for PDGF itself, consistent with the conclusion that the monoclonals recognized only one of two or several receptors for PDGF. Clustering and down-regulation were not seen when the cells were incubated with monovalent Fab' fragments of the PDGFR-B2 antibody. The antibodies also stimulated autophosphorylation of pure PDGF receptor, and PDGFR-B2 was shown to stimulate phosphorylation of phosphofructokinase, an exogenous substrate for the PDGF receptor kinase. High concentrations of PDGFR-B2 antibody, or Fab' fragments thereof, failed to enhance the PDGF receptor kinase activity, compatible with the possibility that dimerization was of importance in the antibody-stimulated kinase activity of purified PDGF receptors.},
  author       = {Rönnstrand, Lars and Terracio, Louis and Claesson-Welsh, Lena and Heldin, Carl-Henrik and Rubin, Kristofer},
  issn         = {1083-351X},
  keyword      = {Animals
*Antibodies,Cell Surface/*immunology/metabolism
Receptors,Monoclonal
Antigen-Antibody Complex
Enzyme-Linked Immunosorbent Assay
Epitopes/*analysis
Female
Kinetics
Platelet-Derived Growth Factor/metabolism
Receptors,Platelet-Derived Growth Factor
Swine
Uterus/metabolism},
  language     = {eng},
  number       = {21},
  pages        = {10429--10435},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {Characterization of two monoclonal antibodies reactive with the external domain of the platelet-derived growth factor receptor},
  volume       = {263},
  year         = {1988},
}