Figure 3.
Fig. 3. C-terminal truncations of endosialidases.
Schematic representations of wild type (wt) and C-terminal
truncated forms are shown in A and B for endoNE and endoNF,
respectively. The positions of the identified cleavage site are
indicated by arrowheads. The C-terminal amino acid and the
number of deleted ( ) amino
acids are shown. In the case of the C-terminal fragment of
endoNE the numbers of the first and last amino acid are shown.
C, soluble fractions of E. coli BL21(DE3) expressing
C-terminally His[6]-tagged wild type or C-terminally truncated
constructs of endoNE were analyzed by 12% ProSieve SDS-PAGE and
immunoblotting. A polyclonal anti-endoNE guinea pig serum was
used for detection. A faint 75 kDa band that is also visible in
the first lane showing lysate of mock transformed bacteria is
visualized by cross-reactivity of the polyclonal serum. Bands
corresponding to full-length and cleaved wild type endoNE are
indicated with arrows. D, wild type and C-terminally truncated
endoNF containing an N-terminal T7 tag were analyzed by 10%
SDS-PAGE and immunoblotting using an anti-T7 antibody.
) amino
acids are shown. In the case of the C-terminal fragment of
endoNE the numbers of the first and last amino acid are shown.
C, soluble fractions of E. coli BL21(DE3) expressing
C-terminally His[6]-tagged wild type or C-terminally truncated
constructs of endoNE were analyzed by 12% ProSieve SDS-PAGE and
immunoblotting. A polyclonal anti-endoNE guinea pig serum was
used for detection. A faint 75 kDa band that is also visible in
the first lane showing lysate of mock transformed bacteria is
visualized by cross-reactivity of the polyclonal serum. Bands
corresponding to full-length and cleaved wild type endoNE are
indicated with arrows. D, wild type and C-terminally truncated
endoNF containing an N-terminal T7 tag were analyzed by 10%
SDS-PAGE and immunoblotting using an anti-T7 antibody.