Biology Reference
In-Depth Information
13.1.3.5.2
Analysis
Data are analyzed as described previously (
Ross & Dixit, 2010
).
DISCUSSION AND SUMMARY
Obtaining active enzyme from every purification is a great challenge. Purifying kata-
nin has proved to be a hard task as inferred by the dearth of publications that under-
take their biochemical and biophysical characterization. The lack of stability is
common in some other AAA
þ
proteins since dynein and CLPX are also known
to lose activity.
The protocol and the assays explained here provide the first steps in the process of
further understanding the biochemical and biophysical properties of microtubule-
severing enzymes. These techniques will be useful in clarifying the role that this
novel class of enzymes has on regulating microtubules.
Acknowledgments
We would like to thank David Sharp, Dong “Ray” Zhang, and Suranjana Mukherjee for their
collaboration, advise, and initial preparation of the bacmid and virus for the Drosophila
GFP-katanin. This work was supported by March of Dimes Basil O'Connor Starter Grant
#5-FY09-46 and NSF CMMI-0928540 to J. L. R. J. L. R. was also supported by NSF MRI
Grant DBI-0923318, and a Cottrell Scholars Award from Research Corporation for Science
Advancement #20031. M. B. and J. D. D.-V. were supported by NSF Grant DMR-1207783.
References
