Heparin-like glycosaminoglycans influence growth and phenotype of human arterial smooth muscle cells in vitro. II. The platelet-derived growth factor A-chain contains a sequence that specifically binds heparin.
Synthetic oligopeptides were used to study the specificity of the interaction between heparin and platelet-derived growth factor (PDGF) in competition experiments. DNA synthesis in PDGF-dependent human arterial smooth muscle cell (hASMC) cultures was used as a biological tracer of PDGF activity. Oligo-108-124 (corresponding to amino acid residues 108-124 of the long PDGF A-chain isoform) had no effect on DNA synthesis in itself but competed at 10(-10) M concentration effectively with PDGF for binding to heparin and released the block on thymidine incorporation induced by heparin. Poly-lysine-serine (lysine:serine ratio 3:1) was also effective but at a considerably higher concentration (10(-6) M). Poly-arginine-serine did not compete with PDGF for heparin as deduced from the cell assay. This suggested that among basic amino acids, lysine was more important than arginine for heparin binding. Deletion of lysine residues 115 and 116 in Oligo-108-124 abolished its effect on the interaction between PDGF and heparin in the cell assay. Likewise, Oligo-69-84 (corresponding to the PDGF A-chain residues 69-84), with three lysine residues interrupted by a proline, was ineffective. In Oligo-108-124, the lysine residues are interrupted by an arginine. Our results suggested that the binding between PDGF and heparin is specific and that the amino acid sequence [-Lys115-Lys116-Arg117-Lys118-Arg119-] is of major importance. They do not however, exclude other domains of the PDGF A or B chains as additional binding sites for heparin nor do they exclude the possibility that heparin and the PDGF receptor share a common binding site.
Fager
G
G
Wallenberg Laboratory for Cardiovascular Research, Faculty of Medicine, University of Göteborg, Sweden.
Camejo
G
G
Olsson
U
U
Ostergren-Lundén
G
G
Bondjers
G
G
D000595:Amino Acid Sequence; D001665:Binding Sites; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D003470:Culture Media; D004247:DNA / Q000096:biosynthesis; D006493:Heparin / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000378:metabolism; D009842:Oligopeptides / Q000378:metabolism; D010982:Platelet-Derived Growth Factor / Q000737:chemistry / Q000378:metabolism*; D012685:Sepharose / Q000031:analogs & derivatives
D003470:Culture Media; D009842:Oligopeptides; D010982:Platelet-Derived Growth Factor; C037405:heparin-sepharose; D006493:Heparin; D004247:DNA; D012685:Sepharose