Predicting the binding properties of cibacron blue F3GA in affinity separation systems
[ X ]
Tarih
2007
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Elsevier
Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
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.
Açıklama
Anahtar Kelimeler
Cibacron Blue F3ga, Quinone Reductase, Hsa, Molecular Dynamics, Affinity, Amber, Mm-Pbsa, Electrostatic Surface Map
Kaynak
International Journal of Biological Macromolecules
WoS Q Değeri
Q4
Scopus Q Değeri
Q1
Cilt
41
Sayı
4
