Nk. Velu et al., REFOLDING OF RIBOFLAVIN CARRIER PROTEIN AS PROBED BY BIOCHEMICAL AND IMMUNOLOGICAL PARAMETERS, Biochimica et biophysica acta. Protein structure and molecular enzymology, 1293(2), 1996, pp. 231-237
The unfolding of the chicken egg white riboflavin carrier protein by d
isulfide reduction with dithiothreitol led to aggregation with concomi
tant loss of ligand binding characteristics and the capacity to intera
ct with six monoclonal antibodies directed against surface-exposed dis
continuous epitopes. The reduced protein could, however, bind to a mon
oclonal antibody recognizing sequential epitope. Under optimal conditi
ons of protein refolding, the vitamin carrier protein regained its fol
ded structure with high efficiency with simultaneous complete restorat
ion of hydrophobic flavin binding site as well as the epitopic conform
ations exposed at the surface in a manner comparable to its native for
m.