title: Electrostatic Forces as Dominant Interactions Between Proteins and Polyanions: an ESI MS Study of Fibroblast Growth Factor Binding to Heparin Oligomers.
abstract: The interactions between fibroblast growth factors (FGFs) and their receptors (FGFRs) are facilitated by heparan sulfate (HS) and heparin (Hp), highly sulfated biological polyelectrolytes. The molecular basis of FGF interactions with these polyelectrolytes is highly complex due to the structural heterogeneity of HS/Hp, and many details still remain elusive, especially the significance of charge density and minimal chain length of HS/Hp in growth factor recognition and multimerization. In this work, we use electrospray ionization mass spectrometry (ESI MS) to investigate the association of relatively homogeneous oligoheparins (octamer, dp8, and decamer, dp10) with acidic fibroblast growth factor (FGF-1). This growth factor forms 1:1, 2:1, and 3:1 protein/heparinoid complexes with both dp8 and dp10, and the fraction of bound protein is highly dependent on protein/heparinoid molar ratio. Multimeric complexes are preferentially formed on the highly sulfated Hp oligomers. Although a variety of oligomers appear to be binding-competent, there is a strong correlation between the affinity and the overall level of sulfation (the highest charge density polyanions binding FGF most strongly via multivalent interactions). These results show that the interactions between FGF-1 and Hp oligomers are primarily directed by electrostatics, and also demonstrate the power of ESI MS as a tool to study multiple binding equilibria between proteins and structurally heterogeneous polyanions. Graphical Abstract ᅟ.
authors.affiliation: Department of Chemistry, University of Massachusetts-Amherst, Amherst, MA, 01003, USA.
authors.forename: Burcu Baykal
authors.initials: BB
authors.lastname: Minsky
authors.affiliation: Department of Chemistry, University of Massachusetts-Amherst, Amherst, MA, 01003, USA.
authors.forename: Paul L
authors.initials: PL
authors.lastname: Dubin
authors.affiliation: Department of Chemistry, University of Massachusetts-Amherst, Amherst, MA, 01003, USA. kaltashov@chem.umass.edu.
authors.forename: Igor A
authors.identifier: 0000-0002-4355-6039
authors.initials: IA
authors.lastname: Kaltashov
mesh_terms: D016220:Fibroblast Growth Factor 1 / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D000071228:Polyelectrolytes; D011108:Polymers / Q000378:metabolism*; D011485:Protein Binding; D055503:Protein Multimerization; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D055672:Static Electricity
chemical_list: D016220:Fibroblast Growth Factor 1; D006493:Heparin; D000071228:Polyelectrolytes; D011108:Polymers
keywords: Glycosaminoglycans; Heparin; Native electrospray ionization mass spectrometry; Noncovalent interactions; Polyelectrolytes