Quantification of Microtubule-Bundling Activity of MAPs Using TIRF Microscopy

Sharol Schmidt-Marcec; Austin Ross; Andrei Smertenko

doi:10.1007/978-1-0716-2867-6_1

Quantification of Microtubule-Bundling Activity of MAPs Using TIRF Microscopy

Methods Mol Biol. 2023:2604:1-12. doi: 10.1007/978-1-0716-2867-6_1.

Authors

Sharol Schmidt-Marcec¹, Austin Ross¹, Andrei Smertenko²

Affiliations

¹ Institute of Biological Chemistry, Washington State University, Pullman, WA, USA.
² Institute of Biological Chemistry, Washington State University, Pullman, WA, USA. andrei.smertenko@wsu.edu.

PMID: 36773221
DOI: 10.1007/978-1-0716-2867-6_1

Abstract

Cross-linking of microtubules by microtubule-associated proteins (MAPs) results in the formation of microtubule bundles. It has been shown that a majority of microtubules in interphase plant cells are bundled. Bundling can contribute to maintaining structural stability and sustaining spatial organization of microtubule arrays. While bundling can be readily detected by an electron or fluorescent microscope, quantifying this activity remains technically challenging. Here we describe a method for quantifying microtubule-bundling in vitro using green and red stable microtubules. Furthermore, this method distinguishes between different types of microtubule-microtubule interactions: bundling, annealing, and branching. Our technique can be used to compare bundling activity of different MAPs and generate parameters for modeling their contribution to organization and dynamics of microtubule arrays.

Keywords: Bundling; Microtubule-associated protein; Quantification; TIRF microscopy.

Publication types

Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Cytoskeleton / metabolism
Microscopy*
Microtubule-Associated Proteins* / metabolism
Microtubules / metabolism

Substances

Microtubule-Associated Proteins

Grants and funding

T32 GM008336/GM/NIGMS NIH HHS/United States