Reconstitution of Neuronal Motor Traffic on Septin-Associated Microtubules

Methods Mol Biol. 2024;2794:79-94. doi: 10.1007/978-1-0716-3810-1_8.ABSTRACTReconstitution of intracellular transport in cell-free in vitro assays enables the understanding and dissection of the molecular mechanisms that underlie membrane traffic. Using total internal reflection fluorescence (TIRF) microscopy and microtubules, which are immobilized to a functionalized glass surface, the kinetic properties of single kinesin molecules can be imaged and analyzed in the presence or absence of microtubule-associated proteins. Here, we describe methods for the in vitro reconstitution of the motility of the neuronal kinesin motor KIF1A on microtubules associated with heteromeric septin (SEPT2/6/7) complexes. This method can be adapted for various neuronal septin complexes and kinesin motors, leading to new insights into the spatial regulation of neuronal membrane traffic by microtubule-associated septins.PMID:38630222 | DOI:10.1007/978-1-0716-3810-1_8
Source: Mol Biol Cell - Category: Molecular Biology Authors: Source Type: research