title: Prolongation of the active lifetime of a biomolecular motor for in vitro motility assay by using an inert atmosphere.
abstract: Over the last few decades, the in vitro motility assay has been performed to probe the biophysical and chemo-mechanical properties as well as the self-organization process of biomolecular motor systems such as actin-myosin and microtubule-kinesin. However, aggression of the reactive oxygen species (ROS) and concomitant termination of the activity of biomolecular motors during investigation remains a drawback of this assay. Despite enzymatic protection that makes use of a combination of glucose, glucose oxidase, and catalase, the active lifetime of biomolecular motors is found to be only a few hours and this short lifetime restricts further study on those systems. We have solved this problem by using a newly developed system of the in vitro motility assay that is conducted in an inert nitrogen gas atmosphere free of ROS. Using microtubule-kinesin as a model system we have shown that our system has prolonged the active lifetime of the biomolecular motor until several days and even a week by protecting it from oxidative damage.
authors.affiliation: Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo 060-0810, Japan.
authors.forename: Arif Md Rashedul
authors.initials: AM
authors.lastname: Kabir
authors.forename: Daisuke
authors.initials: D
authors.lastname: Inoue
authors.forename: Akira
authors.initials: A
authors.lastname: Kakugo
authors.forename: Akiko
authors.initials: A
authors.lastname: Kamei
authors.forename: Jian Ping
authors.initials: JP
authors.lastname: Gong
mesh_terms: D000818:Animals; D066298:In Vitro Techniques; D020409:Molecular Motor Proteins / Q000737:chemistry*; D010100:Oxygen / Q000032:analysis; D010777:Photochemistry; D013552:Swine
chemical_list: D020409:Molecular Motor Proteins; D010100:Oxygen