Differential binding of platelet-derived growth factor isoforms to glycosaminoglycans.
The platelet-derived growth factor (PDGF) family comprises disulfide-bonded dimeric isoforms and plays a key role in the proliferation and migration of mesenchymal cells. Traditionally, it consists of homo- and heterodimers of A and B polypeptide chains that occur as long (A(L) and B(L)) or short (A(S) and B(S)) isoforms. Short isoforms lack the basic C-terminal extension that mediates binding to heparin. In the present study, we show that certain PDGF isoforms bind in a specific manner to glycosaminoglycans (GAGs). Experiments performed with wild-type and mutant Chinese hamster ovary cells deficient in the synthesis of GAGs revealed that PDGF long isoforms bind to heparan sulfate and chondroitin sulfate, while PDGF short isoforms only bind to heparan sulfate. This was confirmed by digestion of cell surface GAGs with heparitinase and chondroitinase ABC and by incubation with sodium chloride to prevent GAG sulfation. Furthermore, exogenous GAGs inhibited the binding of long isoforms to the cell membrane more efficiently than that of short isoforms. Additionally, we performed surface plasmon resonance experiments to study the inhibition of PDGF isoforms binding to low molecular weight heparin by GAGs. These experiments showed that PDGF-AA(L) and PDGF-BB(S) isoforms bound to GAGs with the highest affinity. In conclusion, PDGF activity at the cell surface may depend on the expression of various cellular GAG species.
García-Olivas
Raquel
R
Department of Cellular Biology, Faculty of Biology, University of Barcelona, Avenida Diagonal 645, 08028, Barcelona, Spain.
Hoebeke
Johan
J
Castel
Susanna
S
Reina
Manuel
M
Fager
Gunnar
G
Lustig
Florentina
F
Vilaró
Senén
S
D000818:Animals; D000918:Antibody Specificity; D000077214:Becaplermin; D016466:CHO Cells; D002809:Chondroitin Sulfates / Q000235:genetics / Q000378:metabolism; D006224:Cricetinae; D003871:Dermatan Sulfate / Q000235:genetics / Q000378:metabolism; D006025:Glycosaminoglycans / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D006497:Heparan Sulfate / Q000235:genetics / Q000378:metabolism; D007536:Isomerism; D008071:Lipoprotein Lipase / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D016296:Mutagenesis; D010982:Platelet-Derived Growth Factor / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects / Q000502:physiology; D020574:Proto-Oncogene Proteins c-sis
D000077214:Becaplermin; D002809:Chondroitin Sulfates; D003871:Dermatan Sulfate; D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D008071:Lipoprotein Lipase; D008565:Membrane Proteins; D010982:Platelet-Derived Growth Factor; D020574:Proto-Oncogene Proteins c-sis; C075278:platelet-derived growth factor A