title: Structure-activity relationships for the binding of polymyxins with human α-1-acid glycoprotein.
abstract: Here, for the first time, we have characterized binding properties of the polymyxin class of antibiotics for human α-1-acid glycoprotein (AGP) using a combination of biophysical techniques. The binding affinity of colistin, polymyxin B, polymyxin B(3), colistin methansulfonate, and colistin nona-peptide was determined by isothermal titration calorimetry (ITC), surface plasma resonance (SPR) and fluorometric assay methods. All assay techniques indicated colistin, polymyxin B and polymyxin B(3) display a moderate binding affinity for AGP. ITC and SPR showed there was no detectable binding affinity for colistin methansulfonate and colistin nona-peptide, suggesting both the positive charges of the diaminobutyric acid (Dab) side chains and the N-terminal fatty acyl chain of the polymyxin molecule are required to drive binding to AGP. In addition, the ITC and fluorometric data suggested that endogenous lipidic substances bound to AGP provide part of the polymyxin binding surface. A molecular model of the polymyxin B(3)-AGP F1*S complex was presented that illustrates the pivotal role of the N-terminal fatty acyl chain and the D-Phe6-L-Leu7 hydrophobic motif of polymyxin B(3) for binding to the cleft-like ligand binding cavity of AGP F1*S variant. The model conforms with the entropy driven binding interaction characterized by ITC which suggests hydrophobic interactions coupled to desolvation events and conformational changes are the primary driving force for polymyxins binding to AGP. Collectively, the data are consistent with a role of this acute-phase reactant protein in the transport of polymyxins in plasma.
authors.affiliation: Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Parkville, Victoria 3052, Australia.
authors.forename: Mohammad A K
authors.initials: MA
authors.lastname: Azad
authors.forename: Johnny X
authors.initials: JX
authors.lastname: Huang
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Cooper
authors.forename: Kade D
authors.initials: KD
authors.lastname: Roberts
authors.forename: Philip E
authors.initials: PE
authors.lastname: Thompson
authors.forename: Roger L
authors.initials: RL
authors.lastname: Nation
authors.forename: Jian
authors.initials: J
authors.lastname: Li
authors.forename: Tony
authors.initials: T
authors.lastname: Velkov
mesh_terms: D006801:Humans; D009961:Orosomucoid / Q000737:chemistry* / Q000378:metabolism*; D011112:Polymyxin B / Q000737:chemistry* / Q000145:classification / Q000378:metabolism*; D011113:Polymyxins / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding / Q000502:physiology; D013329:Structure-Activity Relationship
chemical_list: D009961:Orosomucoid; D011113:Polymyxins; D011112:Polymyxin B