Structural and immunological characterisation of partially reduced chickenriboflavin carrier protein

To improve the accessibility of highly folded chicken riboflavin carrier pr otein (cRCP) towards proteolytic enzymes, partial disulphide reduction of t he protein was carried out. The structural and immunological alteration fol lowing reduction and alkylation of two out of the total nine disulphide bon ds (2S-CMRCP) of cRCP was studied. The ability of the protein to interact w ith riboflavin was completely abolished with no change in its far UV, CD sp ectra. Monoclonal antibodies directed towards the assembled epitopes of cRC P also interacted with 2S-CMRCP to a similar extent. However, sandwich assa ys with the mAbs revealed that the topography of the epitopes of 2S-CMRCP i s subtly altered. 2S-CMRCP was more accessible to chymotrypsin and two pept ides were obtained by chymotryptic digestion of 2S-CMRCP.