Abstract
The glutaraldehyde fixation method for fixing tissues is attractive for its ease of use and straightforward surface chemistry. We investigated the effect of glutaraldehyde-induced microtubule immobilization on kinesin-driven cargo transport along microtubules and found that at low glutaraldehyde concentrations, the microtubule–kinesin interaction remains unperturbed. Such findings may facilitate the application of the glutaraldehyde fixation method for many in vitro studies aiming to build nanodevices powered by the microtubule–motor protein interaction.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Rent or buy this article
Get just this article for as long as you need it
$39.95
Prices may be subject to local taxes which are calculated during checkout





References
Silva CJSM, Sousa F, Gübitz, G & Cavaco-Paulo A. Chemical modifications on proteins using glutaraldehyde. Food Technol. Biotechnol. 2004;42:51–6.
Bullock GR. The current status of fixation for electron microscopy: a review. J Microsc. 1984;133:1–15.
Betancor L, López-Gallego F, Alonso-Morales N, Dellamora G, Mateo C, Fernandez-Lafuente R, et al. Glutaraldehyde in protein immobilization. In: Guisan, JM editor. Immobilization of enzymes and cells. Humana Press US; 2006. p. 57–64. https://doi.org/10.1007/978-1-59745-053-9_5.
Quiocho FA, Richards FM. The enzymic behavior of carboxypeptidase-A in the solid state*. Biochemistry. 1966;5:4062–76.
Clancy BE, Behnke-Parks WM, Andreasson JOL, Rosenfeld SS, Block SM. A universal pathway for kinesin stepping. Nat Struct Mol Biol. 2011;18:1020–7.
Kakugo A, Sugimoto S, Gong JP, Osada Y. Gel machines constructed from chemically cross-linked actins and myosins. Adv Mater. 2002;14:1124–6.
Kakugo A, Sugimoto S, Shikinaka K, Gong JP, Osada Y. Characteristics of chemically cross-linked myosin gels. J Biomater Sci Polym Ed. 2005;16:203–18.
Kitamura K, Tokunaga M, Iwane AH, Yanagida T. A single myosin head moves along an actin filament with regular steps of 5.3 nanometres. Nature. 1999;397:129–34.
Hess H, Bachand GD, Vogel V. Powering nanodevices with biomolecular motors. Chemistry. 2004;10:2110–6.
Kabir AMR, Inoue D, Hamano Y, Mayama H, Sada K, Kakugo A. Biomolecular motor modulates mechanical property of microtubule. Biomacromolecules. 2014;15:1797–805.
Inoue D, Nitta T, Kabir AMR, Sada K, Gong JP, Konagaya A, et al. Sensing surface mechanical deformation using active probes driven by motor proteins. Nat Commun. 2016;7:12557.
Keya JJ, Suzuki R, Kabir AMR, Inoue D, Asanuma H, Sada K, et al. DNA-assisted swarm control in a biomolecular motor system. Nat Commun. 2018;9:4–11.
Matsuda K, Kabir AMR, Akamatsu N, Saito A, Ishikawa S, Matsuyama T, et al. Artificial smooth muscle model composed of hierarchically ordered microtubule asters mediated by DNA origami nanostructures. Nano Lett. 2019;19:3933–8.
Sucran Seker, YME in Nanopatterning Nanoscale Devices Biol. Appl. (ed. Seila Šelimović) 261–90 CRC Press, Taylor and Francis Group, 2017.
Nasrin SR, Rashedul Kabir AM, Konagaya A, Ishihara T, Sada K, Kakugo A. Stabilization of microtubules by cevipabulin. Biochem Biophys Res Commun. 2019;516:760-4. https://doi.org/10.1016/j.bbrc.2019.06.095.
Turner D, Chang C, Fang K, Cuomo P, Murphy D. Kinesin movement on glutaraldehyde-fixed microtubules. Anal Biochem. 1996;242:20–5.
Castoldi M, Popov AV. Purification of brain tubulin through two cycles of polymerization-depolymerization in a high-molarity buffer. Protein Expr Purif. 2003;32:83–8.
Peloquin J, Komarova Y, Borisy G. Conjugation of fluorophores to tubulin. Nat Methods. 2005;2:299–303.
Fujimoto K, Kitamura M, Yokokawa M, Kanno I, Kotera H, Yokokawa R. Colocalization of quantum dots by reactive molecules carried by motor proteins on polarized microtubule arrays. ACS Nano. 2013;7:447–55.
Sikora A, Canova FF, Kim K, Nakazawa H, Umetsu M, Kumagai I, et al. Behavior of kinesin driven quantum dots trapped in a microtubule loop. ACS Nano. 2015;9:11003–13.
Conway L, Wood D, Tuzel E, Ross JL. Motor transport of self-assembled cargos in crowded environments. Proc Natl Acad Sci USA. 2012;109:20814–9.
Fernandez-Lafuente R, Rosell CM, Rodriguez V, Guisan JM. Strategies for enzyme stabilization by intramolecular crosslinking with bifunctional reagents. Enzym Microb Technol. 1995;17:517–23.
Acknowledgements
This work was financially supported by the Future AI and Robot Technology Research and Development Project from New Energy and Industrial Technology Development Organization (NEDO), Japan, Grant-in-Aid for Scientific Research on Innovative Areas “Molecular Engine” (JP18H05423) and Grant-in-Aid for Scientific Research (A) (JP18H03673) to AK from the Japan Society for the Promotion of Science (JSPS).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Nasrin, S.R., Kabir, A.M.R., Sada, K. et al. Effect of microtubule immobilization by glutaraldehyde on kinesin-driven cargo transport. Polym J 52, 655–660 (2020). https://doi.org/10.1038/s41428-020-0309-x
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/s41428-020-0309-x