Equilibrium denaturation studies of mouse beta-nerve growth factor.

摘要: Equilibrium denaturation of dimeric mouse beta-nerve growth factor (beta-NGF) has been studied by monitoring changes in the protein's spectroscopic characteristics. Denaturation beta-NGF guanidine hydrochloride and urea resulted an altered intrinsic fluorescence emission spectrum, depolarization, diminished negative circular dichroism. Native-like properties specific biological activity are restored when denaturant is diluted from unfolded samples, demonstrating that this process fully reversible. However, refolding denatured dependent on three disulfide bonds present native protein does not readily occur reduced. Graphical analysis nonlinear least-squares fitting data demonstrate concentration consistent with a two-state model involving dimer monomer (N2 = 2D). The conformational stability calculated according to 19.3 +/- 1.1 kcal/mol 100 mM sodium phosphate at pH 7. Increasing hydrogen ion 25% decrease 4 relative