Ch-AOMK labels C. perfringens BSH expressed
in Escherichia coli. Ch-AOMK (500 μM) was incubated
with
lysates from C. perfringens wildtype (WT) BSH or
C2S mutant expressed in E. coli at 37 °C for
24 h. Following Ch-AOMK labeling, CuAAC tagging was carried out with
(a) Fluor 488-alkyne or (b) biotin-alkyne. (a) Samples were subjected
to SDS-PAGE, and the gel was visualized using fluorescence, followed
by Coomassie staining. (b) Samples were analyzed either by Western
blot with streptavidin-HRP or by silver staining. Input is 2% of the
elution. The arrowhead indicates the expected size of BSH (37 kDa).
A.U. = arbitrary unit. The bands were quantified by densitometry using
ImageJ (bottom panels). Error bars represent standard deviation from
the mean. * p < 0.05, ** p <
0.01, *** p < 0.001, n.s. = not significant, n = (a) 6, (b) 5.