mohammad tabish ahmed - eTheses Repository - University of ...

Chapter 5
In this experiment GroEL was the positive control, while bovine serum albumin (BSA) was
used to measure the background reading. Cpn60.2 was also included as a comparison for the
JNL ATPase activity. The background activity obtained from BSA would be used to correct
the observed activity of the chaperonins. In each case, the OD 620 reading was taken after 30
minutes of incubation with the gold mix.
The results that were obtained showed that the ATPase activity of JNL and Cpn60.2 changed
considerably depending on the buffer used. When compared with the activity of GroEL it
becomes apparent that the activity of JNL and Cpn60.2 is much lower in buffer 5 than it is in
buffer A. Once corrected for the background activity, if GroEL activity is taken as 100%,
then the activity of JNL in buffer 5 is almost nonexistent at just 0.49 ± 2.49% of GroEL (fig
5.8). As expected the ATPase activity of Cpn60.2 in buffer 5 is also extremely low at just
8.27 ± 3.82% of GroEL. In buffer A however, both JNL and Cpn60.2 have higher ATPase
activity, although still not as high as GroEL. Both Cpn60.2 and JNL are at approximately
32.19 ± 1.66% and 25.86 ± 9.42% of GroEL respectively (fig 5.9). The results of these
experiments support the original hypothesis that in conditions where oligomerisation can
occur, the ATPase activity of Cpn60.2 and JNL increases.
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