Immunological crossreactivity of Paramecium surface antigens: conformational differences account for cell surface specificity.

Abstract

Surface antigens (i-Ags) G and D of P. primaurelia, reported earlier to be completely unrelated on the basis of peptide mapping and immunological tests, were compared and found to be similar in MW (235,000), isoelectric point (.apprx. 4.5) and amino acid composition. They are tightly folded proteins with > 100 intramolecular disulfide bonds. Native i-Ags and their CNBr fragments were blotted onto nitrocellulose and labeled with a battery of homologous and heterologous antibodies fractionated on native and reduced i-Ag immunoadsorbents. The i-Ags retained part of their antigenic structure upon cleavage and reduction. Analysis of labeling revealed that the 2 proteins share many of their antigenic sites, though they are completely distinct in their native conformation. Immunofluorescence and immunoadsorption on fixed cells confirmed these results. Antigenic crossreactivity also was found with surface antigens of a ciliate outside the Paramecium spp. complex. Surface specificity of paramecia is due to the expression of related proteins in different conformations.