Refolding of riboflavin carrier protein as probed by biochemical and immunological parameters.
The unfolding of the chicken egg white riboflavin carrier protein by disulfide reduction with dithiothreitol led to aggregation with concomitant loss of ligand binding characteristics and the capacity to interact with six monoclonal antibodies directed against surface-exposed discontinuous epitopes. The reduced protein could, however, bind to a monoclonal antibody recognizing sequential epitope. Under optimal conditions of protein refolding, the vitamin carrier protein regained its folded structure with high efficiency with simultaneous complete restoration of hydrophobic flavin binding site as well as the epitopic conformations exposed at the surface in a manner comparable to its native form.
Velu
N K
NK
Centre for Reproductive Biology and Molecular Endocrinology, Indian Institute of Science, Bangalore, India.
Karande
A A
AA
Adiga
P R
PR
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002352:Carrier Proteins / Q000737:chemistry* / Q000276:immunology; D002645:Chickens; D002850:Chromatography, Gel; D004220:Disulfides / Q000737:chemistry; D004229:Dithiothreitol / Q000494:pharmacology; D004529:Egg White; D000939:Epitopes / Q000276:immunology; D019791:Guanidine; D006146:Guanidines; D026901:Membrane Transport Proteins; D010084:Oxidation-Reduction; D011485:Protein Binding; D011489:Protein Denaturation; D017510:Protein Folding; D012256:Riboflavin / Q000378:metabolism*; D013050:Spectrometry, Fluorescence
D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D004220:Disulfides; D000939:Epitopes; D006146:Guanidines; D026901:Membrane Transport Proteins; C019079:riboflavin-binding protein; D019791:Guanidine; D004229:Dithiothreitol; D012256:Riboflavin