Platelet-derived growth factor-BB and basic fibroblast growth factor directly interact in vitro with high affinity

Katia Russo, Raffaele Ragone, Angelo M. Facchiano, Maurizio C. Capogrossi, Antonio Facchiano

Research output: Contribution to journalArticle

Abstract

Platelet-derived growth factor-BB (PDGF-BB) and basic fibroblast growth factor (bFGF) are potent growth factors active on many cell types. The present study indicates that they directly interact in vitro. The interaction was investigated with overlay experiments, surface plasmon resonance experiments, and solid-phase immunoassays by immobilizing one factor or the other and by steady-state fluorescence analysis. The interaction observed was specific, dose-dependent, and saturable, and the bFGF/PDGF-BB binding stoichiometry was found to be 2:1. K D1 for the first step equilibrium and the overall K D values were found to be in the nanomolar and in the picomolar range, respectively. Basic FGF/PDGF-BB interaction was strongly reduced as a function of time of PDGF-BB proteolysis. Furthermore, docking analysis suggested that the PDGF-BB region interacting with bFGF may overlap, at least in part, with the PDGF-BB receptor-binding site. This hypothesis was supported by surface plasmon resonance experiments showing that an anti-PDGF-BB antibody, known to inhibit PDGF-BB binding with its receptor, strongly reduced bFGF/PDGF-BB interaction, whereas a control antibody was ineffective. According to these data, the observed bFGF-PDGF-BB complex formation might explain, at least in part, previous observations showing that PDGF-BB chemotactic and mitogenic activity on smooth muscle cells are strongly inhibited in the presence of bFGF.

Original languageEnglish
Pages (from-to)1284-1291
Number of pages8
JournalJournal of Biological Chemistry
Volume277
Issue number2
DOIs
Publication statusPublished - Jan 11 2002

Fingerprint

Fibroblast Growth Factor 2
Surface Plasmon Resonance
Surface plasmon resonance
platelet-derived growth factor BB
In Vitro Techniques
Proteolysis
Platelet-Derived Growth Factor Receptors
Antibodies
Experiments
Immunoassay
Stoichiometry
Smooth Muscle Myocytes
Muscle
Intercellular Signaling Peptides and Proteins
Fluorescence
Binding Sites
Cells

ASJC Scopus subject areas

  • Biochemistry

Cite this

Platelet-derived growth factor-BB and basic fibroblast growth factor directly interact in vitro with high affinity. / Russo, Katia; Ragone, Raffaele; Facchiano, Angelo M.; Capogrossi, Maurizio C.; Facchiano, Antonio.

In: Journal of Biological Chemistry, Vol. 277, No. 2, 11.01.2002, p. 1284-1291.

Research output: Contribution to journalArticle

Russo, Katia ; Ragone, Raffaele ; Facchiano, Angelo M. ; Capogrossi, Maurizio C. ; Facchiano, Antonio. / Platelet-derived growth factor-BB and basic fibroblast growth factor directly interact in vitro with high affinity. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 2. pp. 1284-1291.
@article{3717a1b6922b427687eef1360722d934,
title = "Platelet-derived growth factor-BB and basic fibroblast growth factor directly interact in vitro with high affinity",
abstract = "Platelet-derived growth factor-BB (PDGF-BB) and basic fibroblast growth factor (bFGF) are potent growth factors active on many cell types. The present study indicates that they directly interact in vitro. The interaction was investigated with overlay experiments, surface plasmon resonance experiments, and solid-phase immunoassays by immobilizing one factor or the other and by steady-state fluorescence analysis. The interaction observed was specific, dose-dependent, and saturable, and the bFGF/PDGF-BB binding stoichiometry was found to be 2:1. K D1 for the first step equilibrium and the overall K D values were found to be in the nanomolar and in the picomolar range, respectively. Basic FGF/PDGF-BB interaction was strongly reduced as a function of time of PDGF-BB proteolysis. Furthermore, docking analysis suggested that the PDGF-BB region interacting with bFGF may overlap, at least in part, with the PDGF-BB receptor-binding site. This hypothesis was supported by surface plasmon resonance experiments showing that an anti-PDGF-BB antibody, known to inhibit PDGF-BB binding with its receptor, strongly reduced bFGF/PDGF-BB interaction, whereas a control antibody was ineffective. According to these data, the observed bFGF-PDGF-BB complex formation might explain, at least in part, previous observations showing that PDGF-BB chemotactic and mitogenic activity on smooth muscle cells are strongly inhibited in the presence of bFGF.",
author = "Katia Russo and Raffaele Ragone and Facchiano, {Angelo M.} and Capogrossi, {Maurizio C.} and Antonio Facchiano",
year = "2002",
month = "1",
day = "11",
doi = "10.1074/jbc.M108858200",
language = "English",
volume = "277",
pages = "1284--1291",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "2",

}

TY - JOUR

T1 - Platelet-derived growth factor-BB and basic fibroblast growth factor directly interact in vitro with high affinity

AU - Russo, Katia

AU - Ragone, Raffaele

AU - Facchiano, Angelo M.

AU - Capogrossi, Maurizio C.

AU - Facchiano, Antonio

PY - 2002/1/11

Y1 - 2002/1/11

N2 - Platelet-derived growth factor-BB (PDGF-BB) and basic fibroblast growth factor (bFGF) are potent growth factors active on many cell types. The present study indicates that they directly interact in vitro. The interaction was investigated with overlay experiments, surface plasmon resonance experiments, and solid-phase immunoassays by immobilizing one factor or the other and by steady-state fluorescence analysis. The interaction observed was specific, dose-dependent, and saturable, and the bFGF/PDGF-BB binding stoichiometry was found to be 2:1. K D1 for the first step equilibrium and the overall K D values were found to be in the nanomolar and in the picomolar range, respectively. Basic FGF/PDGF-BB interaction was strongly reduced as a function of time of PDGF-BB proteolysis. Furthermore, docking analysis suggested that the PDGF-BB region interacting with bFGF may overlap, at least in part, with the PDGF-BB receptor-binding site. This hypothesis was supported by surface plasmon resonance experiments showing that an anti-PDGF-BB antibody, known to inhibit PDGF-BB binding with its receptor, strongly reduced bFGF/PDGF-BB interaction, whereas a control antibody was ineffective. According to these data, the observed bFGF-PDGF-BB complex formation might explain, at least in part, previous observations showing that PDGF-BB chemotactic and mitogenic activity on smooth muscle cells are strongly inhibited in the presence of bFGF.

AB - Platelet-derived growth factor-BB (PDGF-BB) and basic fibroblast growth factor (bFGF) are potent growth factors active on many cell types. The present study indicates that they directly interact in vitro. The interaction was investigated with overlay experiments, surface plasmon resonance experiments, and solid-phase immunoassays by immobilizing one factor or the other and by steady-state fluorescence analysis. The interaction observed was specific, dose-dependent, and saturable, and the bFGF/PDGF-BB binding stoichiometry was found to be 2:1. K D1 for the first step equilibrium and the overall K D values were found to be in the nanomolar and in the picomolar range, respectively. Basic FGF/PDGF-BB interaction was strongly reduced as a function of time of PDGF-BB proteolysis. Furthermore, docking analysis suggested that the PDGF-BB region interacting with bFGF may overlap, at least in part, with the PDGF-BB receptor-binding site. This hypothesis was supported by surface plasmon resonance experiments showing that an anti-PDGF-BB antibody, known to inhibit PDGF-BB binding with its receptor, strongly reduced bFGF/PDGF-BB interaction, whereas a control antibody was ineffective. According to these data, the observed bFGF-PDGF-BB complex formation might explain, at least in part, previous observations showing that PDGF-BB chemotactic and mitogenic activity on smooth muscle cells are strongly inhibited in the presence of bFGF.

UR - http://www.scopus.com/inward/record.url?scp=0037059794&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037059794&partnerID=8YFLogxK

U2 - 10.1074/jbc.M108858200

DO - 10.1074/jbc.M108858200

M3 - Article

C2 - 11694520

AN - SCOPUS:0037059794

VL - 277

SP - 1284

EP - 1291

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 2

ER -