Akademik Veri Yönetim Sistemi
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, cilt.41, sa.4, ss.430-438, 2007 (SCI-Expanded)
The binding properties of cibacron blue F3GA (CB-F3GA) bound to a model NAD(P)H/FAD(H-2)-dependent protein system, namely cytosolic quinone reductase (QR), was characterized by AMBER in an attempt to address the binding properties of immobilized CB-F3GA used in the separation of serum albumin. A favorable binding free energy of -4.52 kcal/mol (K-D = 5.09 x 10(-4) kcal/mol) was determined for CB-F3GA binding by MM-PBSA method, which was found to be a ballpark estimate of empirical values reported in literature (Delta G approximate to -6 kcal/mol). We propose that CB-F3GA primarily follows a class III binding motif in presence of FAD in the binding site of QR in solution, while a class II binding motif is observed in the crystal form. It was found that favorable van der Waals/hydrophobic interactions take place in the binding site making a major contribution to a favorably dominating enthalpy of binding (Delta H-tot = -25.87 kcal/mol) as compared to a disfavorable binding entropy term (T Delta S-tot = -21.35 kcal/mol). Additional MM-PBSA experiments in the absence of FAD gave rise to a disfavorable binding free energy for CB in complex with QR, suggesting that FAD is an essential determinant of CB-F3GA binding. This is in contrast to an earlier observation of Denizli et al. on separation of human serum albumin (HSA) by immobilized CB-F3GA in the absence of FAD. Therefore, a class I binding model for CB-F3GA is proposed here to account for the efficient separation of HSA in affinity chromatography systems. (C) 2007 Elsevier B.V. All rights reserved.