title: Tug-of-war of microtubule filaments at the boundary of a kinesin- and dynein-patterned surface.
abstract: Intracellular cargo is transported by multiple motor proteins. Because of the force balance of motors with mixed polarities, cargo moves bidirectionally to achieve biological functions. Here, we propose a microtubule gliding assay for a tug-of-war study of kinesin and dynein. A boundary of the two motor groups is created by photolithographically patterning gold to selectively attach kinesin to the glass and dynein to the gold surface using a self-assembled monolayer. The relationship between the ratio of two antagonistic motor numbers and the velocity is derived from a force-velocity relationship for each motor to calculate the detachment force and motor backward velocity. Although the tug-of-war involves >100 motors, values are calculated for a single molecule and reflect the collective dynein and non-collective kinesin functions when they work as a team. This assay would be useful for detailed in vitro analysis of intracellular motility, e.g., mitosis, where a large number of motors with mixed polarities are involved.
authors.affiliation: Department of Micro Engineering, Kyoto University, Kyoto 615-8540, Japan.
authors.forename: Junya
authors.initials: J
authors.lastname: Ikuta
authors.affiliation: Department of Micro Engineering, Kyoto University, Kyoto 615-8540, Japan.
authors.forename: Nagendra K
authors.initials: NK
authors.lastname: Kamisetty
authors.affiliation: Department of Micro Engineering, Kyoto University, Kyoto 615-8540, Japan.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Shintaku
authors.affiliation: Department of Micro Engineering, Kyoto University, Kyoto 615-8540, Japan.
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Kotera
authors.affiliation: 1] Faculty of Bioscience and Applied Chemistry, Department of Frontier Bioscience, Hosei University, Koganei 184-8584, Japan [2] Precursory Research for Embryonic Science and Technology (PRESTO) from the Japan Science and Technology Agency (JST), Kawaguchi, Saitama 332-0012, Japan.
authors.forename: Takahide
authors.initials: T
authors.lastname: Kon
authors.affiliation: 1] Department of Micro Engineering, Kyoto University, Kyoto 615-8540, Japan [2] Precursory Research for Embryonic Science and Technology (PRESTO) from the Japan Science and Technology Agency (JST), Kawaguchi, Saitama 332-0012, Japan.
authors.forename: Ryuji
authors.initials: R
authors.lastname: Yokokawa
mesh_terms: D000465:Algorithms; D001692:Biological Transport; D004398:Dyneins / Q000737:chemistry* / Q000378:metabolism; D016547:Kinesins / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D008870:Microtubules / Q000737:chemistry* / Q000378:metabolism; D008954:Models, Biological; D008958:Models, Molecular; D020409:Molecular Motor Proteins / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D055503:Protein Multimerization; D014743:Videotape Recording
chemical_list: D020409:Molecular Motor Proteins; D004398:Dyneins; D016547:Kinesins