Measuring the Effects of Microtubule-Associated Proteins on Microtubule Dynamics In Vitro

Methods in Molecular Biology
Marija Zanic


Microtubule dynamic instability, the process by which individual microtubules switch between phases of growth and shrinkage, is essential for establishing the architecture of cellular microtubule structures, such as the mitotic spindle. This switching process is regulated by a complex network of microtubule-associated proteins (MAPs), which modulate different aspects of microtubule dynamic behavior. To elucidate the effects of MAPs and their molecular mechanisms of action, in vitro reconstitution approaches with purified components are used. Here, I present methods for measuring individual and combined effects of MAPs on microtubule dynamics, using purified protein components and total-internal-reflection fluorescence (TIRF) microscopy. Particular focus is given to the experimental design, proper parameterization, and data analysis.


Aug 20, 2019·The Journal of Cell Biology·Claire StrothmanMarija Zanic
Mar 9, 2019·Scientific Reports·Varun KapoorSimone Reber
May 28, 2020·Proceedings of the National Academy of Sciences of the United States of America·Göker ArpağMarija Zanic

Related Concepts

Microtubule-Associated Proteins
TUBE1 gene
Microtubule-Associated Protein 3
Mitotic Spindle Apparatus
Time-Lapse Imaging
Immunofluorescence Microscopy
Microtubule Cytoskeleton Organization

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