A. Deshmukh et al., Structural and immunological characterisation of partially reduced chickenriboflavin carrier protein, I J BIOCH B, 37(3), 2000, pp. 166-170
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.