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
- Rönnstrand, Lars
LU
; Terracio, Louis
; Claesson-Welsh, Lena
; Heldin, Carl-Henrik
and Rubin, Kristofer
- publishing date
- 1988
- 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
- American Society for Biochemistry and Molecular Biology
- external identifiers
-
- scopus:0023694016
- ISSN
- 1083-351X
- language
- English
- LU publication?
- no
- additional info
- The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Experimental Clinical Chemistry (013016010)
- id
- 64336f64-e26e-4b85-ac1e-2e8a619bc66f (old id 1784230)
- date added to LUP
- 2016-04-04 07:33:08
- date last changed
- 2021-08-29 03:58:44
@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}},
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}},
language = {{eng}},
number = {{21}},
pages = {{10429--10435}},
publisher = {{American Society for Biochemistry and Molecular Biology}},
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}},
}