title: Importance of pH and disulfide bridges on the structural and binding properties of human α₁-acid glycoprotein.
abstract: Human α(1)-acid glycoprotein (AGP) is an acute phase plasma glycoprotein containing two disulfide bridges. As a member of the lipocalin superfamily, it binds and transports several basic and neutral ligands, but a number of other activities have also been described. Thanks to its binding properties, AGP is also a good candidate for the development of biosensors and affinity chromatography media, and in this context detailed structural information is needed. The structural properties of AGP at different p(2)Hs and under reducing conditions were analysed by FT-IR spectroscopy. The obtained data indicate that AGP, when denatured, does not aggregate at neutral or basic p(2)Hs whilst it does at acidic p(2)Hs. Under reducing conditions the protein is remarkably less thermostable than its oxidized counterpart and presents an enhanced tendency to aggregate, even at neutral p(2)H. A heat-induced molten globule-like state (MG) was detected at 55 °C at p(2)H 7.4 and 5.5. At p(2)H 4.5 the MG occurred at 45 °C with an onset of formation at 40 °C. The MG was not observed under reducing conditions. A lower affinity of chlorpromazine and progesterone for the MG formed at p(2)H 4.5 and 40 °C was observed, suggesting that ligand(s) may be released near the negative surfaces of biological membranes. Furthermore, the reduced AGP displays an enhanced affinity for progesterone, indicating the importance of disulfide bonds for the binding capacity of AGP.
authors.affiliation: Dipartimento di Biochimica Biologia e Genetica, Università Politecnica delle Marche, Ancona, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Scirè
authors.forename: Maurizio
authors.initials: M
authors.lastname: Baldassarre
authors.forename: Giulio
authors.initials: G
authors.lastname: Lupidi
authors.forename: Fabio
authors.initials: F
authors.lastname: Tanfani
mesh_terms: D004220:Disulfides / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008024:Ligands; D009961:Orosomucoid / Q000737:chemistry*; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D013696:Temperature
chemical_list: D004220:Disulfides; D008024:Ligands; D009961:Orosomucoid