title: Acidic pH modulates the interaction between human granulocyte-macrophage colony-stimulating factor and glycosaminoglycans.
abstract: Granulocyte-macrophage colony-stimulating factor (GM-CSF) controls growth and differentiation of hematopoietic cells. Previous reports have indicated that the mitogenic activity of GM-CSF may be modulated by the glycosidic moiety of proteoglycans associated with the membrane of stromal cells. In this work, we have performed in vitro studies of the interaction between GM-CSF and glycosaminoglycans. The addition of heparin promoted a marked blue shift in the fluorescence emission spectrum of GM-CSF as well as a 30-fold increase in the intensity of light scattering, which indicates formation of large molecular weight complexes between the two molecules. Interestingly, heparin-induced changes in the spectral properties of GM-CSF were only observed at acidic pH. The dependence on acidic pH, together with a strict dependence on glycosaminoglycan sulfation and the fact that high ionic strength destabilized the interaction, indicates that the association between GM-CSF and glycosaminoglycans is mediated by electrostatic interactions. These interactions probably involve sulfate groups in the glycosaminoglycans and positively charged histidine residues in GM-CSF. We propose that negatively charged glycolipids present on the plasma membrane of the hematopoietic and/or the stromal cell could promote an acidic microenvironment capable of triggering interaction between GM-CSF and membrane-bound proteoglycans in vivo.
authors.affiliation: Departamento de Bioquímica Médica, Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Cidade Universitária, Rio de Janeiro, RJ 21941-590, Brazil.
authors.forename: A
authors.initials: A
authors.lastname: Wettreich
authors.forename: A
authors.initials: A
authors.lastname: Sebollela
authors.forename: M A
authors.initials: MA
authors.lastname: Carvalho
authors.forename: S P
authors.initials: SP
authors.lastname: Azevedo
authors.forename: R
authors.initials: R
authors.lastname: Borojevic
authors.forename: S T
authors.initials: ST
authors.lastname: Ferreira
authors.forename: T
authors.initials: T
authors.lastname: Coelho-Sampaio
mesh_terms: D001665:Binding Sites; D006025:Glycosaminoglycans / Q000378:metabolism*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008081:Liposomes / Q000378:metabolism; D008958:Models, Molecular; D009282:Naphthalenesulfonates / Q000378:metabolism; D010743:Phospholipids; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism; D012685:Sepharose / Q000031:analogs & derivatives / Q000378:metabolism; D013050:Spectrometry, Fluorescence
chemical_list: D006025:Glycosaminoglycans; D008081:Liposomes; D009282:Naphthalenesulfonates; D010743:Phospholipids; D011994:Recombinant Proteins; C037405:heparin-sepharose; C009497:2-(4-toluidino)-6-naphthalenesulfonic acid; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D006493:Heparin; D012685:Sepharose