1. Purification and biophysical analysis of microtubule-severing enzymes in vitro.
- Author
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Diaz-Valencia JD, Bailey M, and Ross JL
- Subjects
- ATPases Associated with Diverse Cellular Activities, Amino Acid Sequence, Animals, Drosophila Proteins analysis, Drosophila melanogaster, Humans, Katanin, Microtubules metabolism, Molecular Sequence Data, Sf9 Cells cytology, Spastin, Adenosine Triphosphatases analysis, Microtubule-Associated Proteins analysis, Nuclear Proteins analysis
- Abstract
Microtubule-severing enzymes are a novel class of microtubule regulators. They are enzymes from the ATPases associated with various cellular activities family (AAA+) that utilize ATP to cut microtubules into smaller filaments. Discovered over 20 years ago, there are still many open questions about severing enzymes. Both cellular and biochemical studies need to be pursued to fully understand how these enzymes function mechanistically in the cell. Here, we present methods to express, purify, and test the biophysical nature of these proteins in vitro to begin to address the biochemical and biophysical mechanisms of this important and novel group of microtubule destabilizers., (Copyright © 2013 Elsevier Inc. All rights reserved.) more...
- Published
- 2013
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