Characterization and localization of CIV polypeptides.

In order to detect the structural proteins linked with disulfide bonds, CIV was solubilized and electrophoresed under nonreducing conditions in the first dimension and then under reducing conditions in the second dimension. The viral polypeptides linked originally with disulfide bonds were separated into subunits. The complexes were trimers (P'50) or dimers (P60 and P10). The apparent molecular weights of P81, P53, and P49 changed significantly according to the composition of the lysis buffer used, suggesting that the differences in their molecular weights were due to conformational changes produced by reduction of their intramolecular disulfide bonds. Sulfhydryl-containing polypeptides (P'50-P50, P60, P100, and P33) were detected by N-[14C]ethylmaleimide, and the accessibility of these residues was analyzed after successive stripping of the CIV particle. Radioiodination of external polypeptides by [125I]iodosulfanilic acid shows only one intensively labeled spot corresponding to the P50 polypeptide, whereas P'50 was only slightly labeled. Six viral polypeptides P81, P60, P31, P17, P13, and P10 were revealed to possess high affinity for CIV DNA. A structural model of CIV is proposed and discussed.