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Figure 6.
Fig. 6. Trimer formation of endoNF. A, wild type and
mutant endoNF were expressed in E. coli BL21(DE3), and soluble
fractions of the bacterial lysates were analyzed by 6% SDS-PAGE
and Western blot. To visualize SDS-resistant trimers, one
aliquot of each sample was analyzed omitting the boiling step
before electrophoresis. The upper blot was developed with a
combination of anti-T7 and anti-His[6] antibody and the lower
blot with anti-His[6] antibody, exclusively. EndoNF mutants with
the indicated amino acid exchanges are shown in lanes 3-10 and
C-terminal truncated forms in lanes 11-14. Bands corresponding
to trimers, N-terminal fragments, and full-length endoNF are
indicated with arrows. B, wild type endoNF, endoNE, and the
chimera endoNF-E and endoNE-F were monitored for trimer
formation as described above. Bands corresponding to an
SDS-resistant complex and the N-terminal catalytic domains of
endoNF and endoNE are marked with arrows.
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