Life Sci. 1997;61(23):2291-303. Unique Identifier : AIDSLINE
Trichosanthin (TCS) fragments were produced by Tn1000 deletion
mutagenesis and by cyanogen bromide (CNBr) cleavage and their
immunoreactivity was examined by incubating with various antibodies.
Twelve C-terminally truncated TCS variants were successfully synthesized
under the control of a T7 RNA driven promoter. The smallest antigenic
fragment mapped corresponded to the N-terminal 20 amino acids (aa). Six
CNBr fragments of TCS were created and identified by electrospray mass
spectrometry and N-terminal sequencing. Three antigenic fragments
corresponding to aa 1-72, 101-152 and 153-246, respectively were mapped.
Fragments corresponding to aa 1-72 and 153-246 were immunoreactive to
the same monoclonal antibody showing they are components of a
discontinuous epitope. On the other hand, the fragment containing aa
73-100 was not detected by any of the antibodies used.