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However, the mechanism underlying the interactions between kinesin-5 and the microtubule remains poorly understood. In this study, we investigated the binding of BimC, a kinesin-5 motor from Aspergillus nidulans, to the microtubule using a combination of total internal reflection fluorescence (TIRF) microscopy and molecular dynamics (MD) simulations. TIRF microscopy experiments revealed that increasing the concentration of KCl in the motility buffer from 0\u00a0mM to 150\u00a0mM completely abolishes the ability of BimC to bind to the microtubule. Consistent with this experimental finding, MD simulations demonstrated a significant reduction in the strength of electrostatic interactions between BimC and microtubules at 150\u00a0mM KCl compared to 0\u00a0mM KCl. Furthermore, we identified several salt bridges at the BimC-microtubule interface, with positively charged residues on BimC interacting with negatively charged residues on the tubulin heterodimer. These results provide mechanistic insights into the role of electrostatic interactions in kinesin-5\u2013microtubule binding, advancing our understanding of the molecular underpinnings of kinesin-5 motility.<\/jats:p>","DOI":"10.1093\/bib\/bbaf144","type":"journal-article","created":{"date-parts":[[2025,3,18]],"date-time":"2025-03-18T12:43:09Z","timestamp":1742301789000},"source":"Crossref","is-referenced-by-count":4,"title":["Elucidating the interactions between Kinesin-5\/BimC and the microtubule: insights from TIRF microscopy and molecular dynamics simulations"],"prefix":"10.1093","volume":"26","author":[{"given":"Wenhan","family":"Guo","sequence":"first","affiliation":[{"name":"Department of Physics, University of Texas at El Paso , 500 W University Ave, El Paso, TX 79968 ,","place":["United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuan","family":"Gao","sequence":"additional","affiliation":[{"name":"Department of Physics, Oregon State University , 1500 Jefferson Way, Corvallis, OR 97330 ,","place":["United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dan","family":"Du","sequence":"additional","affiliation":[{"name":"Computational Science Program, University of Texas at El Paso , 500 W University Ave, El Paso, TX 79968 ,","place":["United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jason E","family":"Sanchez","sequence":"additional","affiliation":[{"name":"Computational Science Program, University of Texas at El Paso , 500 W University Ave, El Paso, TX 79968 ,","place":["United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yupeng","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, University of Texas at El Paso , 500 W University Ave, El Paso, TX 79968 ,","place":["United States"]},{"name":"Border Biomedical Research Center, University of Texas at El Paso , 500 W University Ave, El Paso, TX 79968 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