TY  - JOUR
TI  - Quantification of Spatial Patterns of Microtubule Transport by Kinesin-1 Head and Tail
AU  - Basu, Jashaswi  
AU  - Singh, Kajal  
AU  - Jannasch, Anita  
AU  - Athale, Chaitanya  A.
VL  - 16
IS  - 10
PY  - 2026
DA  - 2026/05/20
SP  - e5697
C1  - Bio-protocol 2026;16:e5697
DO  - 10.21769/BioProtoc.5697
UR  - https://doi.org/10.21769/BioProtoc.5697
AB  - The conventional kinesin-1 is a plus-end-directed microtubule-dependent motor protein with distinct motor head, stalk, and tail domains. Along with the motor head, which binds and walks along microtubules in an adenosine 5’-triphosphate (ATP) dependent manner, kinesin also contains a C-terminal microtubule binding tail. Motor-driven collective motility is well characterized using in vitro gliding assays, which show uninterrupted, smooth trajectories of transport. However, gliding assays driven by the full-length Drosophila kinesin-1 with both head and tail resulted in the emergence of spontaneous spatial microtubule patterns and stop-and-go motion. This was reproduced by an equimolar ratio of the active head and passive tail. Here, we describe the detailed protocol to reconstitute these microtubule gliding assays using multiple motor types: the full-length kinesin-1, the motor head or tail, mixtures of both head and tail, and a rigor mutant of the kinesin. We provide details of the approach taken to acquire the image time-series, to then quantify the spatial patterns that result from these motor combinations. Our approach provides a framework to systematically characterize the spatiotemporal effects of molecular motor-driven collective microtubule transport.
KW  - Microtubule
KW  - Kinesin-1
KW  - Gliding assay
KW  - Microscopy
KW  - Image analysis
KW  - Gibson assembly
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False