title: Purification and biophysical analysis of microtubule-severing enzymes in vitro.
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.
authors: Department of Physics, University of Massachusetts Amherst, Amherst, Massachusetts, USA.
authors: Juan Daniel
authors: JD
authors: Diaz-Valencia
authors: Megan
authors: M
authors: Bailey
authors: Jennifer L
authors: JL
authors: Ross
mesh_terms: D000074183:ATPases Associated with Diverse Cellular Activities; D000251:Adenosine Triphosphatases / Q000032:analysis*; D000595:Amino Acid Sequence; D000818:Animals; D029721:Drosophila Proteins / Q000032:analysis; D004331:Drosophila melanogaster; D006801:Humans; D000074201:Katanin; D008869:Microtubule-Associated Proteins / Q000032:analysis*; D008870:Microtubules / Q000378:metabolism; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000032:analysis*; D061987:Sf9 Cells / Q000166:cytology; D000074181:Spastin
chemical_list: D029721:Drosophila Proteins; D008869:Microtubule-Associated Proteins; D009687:Nuclear Proteins; D000251:Adenosine Triphosphatases; D000074183:ATPases Associated with Diverse Cellular Activities; C583279:FIGNL1 protein, human; D000074181:Spastin; D000074201:Katanin; C400283:SPAST protein, human
keywords: AAA+; fidgetin; katanin; microtubule destabilizer; severing enzyme; spastin