Polyoxometalate binding to human serum albumin: a thermodynamic and spectroscopic approach.
The molecular recognition of polyoxometalates by human serum albumin is studied using two different polyoxometalates (POMs) at pH 7.5. The results are compared with those obtained at pH 3.5 and 9.0. At pH 7.5, both POMs strongly interact with the protein with different binding behaviors. The Keggin shaped POM, [H(2)W(12)O(40)](6-) (H2W12), specifically binds the protein, forming a complex with a 1:1 stoichiometry with Ka = 2.9 x 10(6) M(-1). The binding constant decreased dramatically with the increase of the ionic strength, thus indicating a mostly electrostatic binding process. Isothermal titration calorimetry (ITC) experiments show that the binding is an enthalpically driven exothermic process. For the wheel shaped POM [NaP(5)W(30)O(110)](14-) (P5W30), there are up to five binding sites on the protein. Increasing the ionic strength changes the binding behavior significantly, leading to a simple exothermic process, with several binding sites. Competitive binding experiments indicate that the two POMs share one common binding site. In addition, they show the existence of another important binding site for P5W30. The two POMs exhibit different binding dependences on the pH. The combination of the experimental results with the knowledge of the surface map of the protein in its N-B conformation transition domain leads to the proposal for the probable binding site of POMs. The present work reveals a protein conformation change upon P5W30 binding, a new feature not explicitly documented in previous studies.
Zhang
Guangjin
G
Laboratoire de Chimie Physique, Equipe d'Electrochimie et Photoélectrochimie, UMR 8000, CNRS, Université Paris-Sud 11, Bâtiment 350, 91405 Orsay Cedex, France.
Keita
Bineta
B
Craescu
Constantin T
CT
Miron
Simona
S
de Oliveira
Pedro
P
Nadjo
Louis
L
D006801:Humans; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D009994:Osmolar Concentration; D011485:Protein Binding; D012709:Serum Albumin / Q000737:chemistry* / Q000378:metabolism*; D013053:Spectrophotometry; D013816:Thermodynamics; D017186:Titrimetry; D017973:Tungsten Compounds / Q000737:chemistry*