Biophysical measurements on axonemal dyneins.
In recent decades, the development of technologies such as optical trap nanometry and advanced fluorescence microscopy have provided tools for studying the dynamics of single protein molecules in vitro and in vivo with nanometer precision over timescales from milliseconds to seconds. The single-molecule sensitivities of these methods permit studies to be made on conformational changes and dynamics of protein molecules that are masked in ensemble-averaged experiments. For protein motors, force generation, processivity, step size, transitions among mechanical states, and mechanochemical coupling are among the properties that can be directly measured by single-molecule techniques. Our understanding of the functions of protein motors has thus benefited considerably from the application of single-molecule techniques. This chapter will focus on single-molecule techniques applicable to axonemal dyneins, the principles upon which they work and how they are constructed and conducted.
Kojima
Hiroaki
H
Kobe Advanced ICT Research Center, National Institute of Information and Communications Technology, Nishi-ku, Japan.
Toba
Shiori
S
Sakakibara
Hitoshi
H
Oiwa
Kazuhiro
K
D000818:Animals; D056444:Axonemal Dyneins / Q000737:chemistry* / Q000378:metabolism; D001681:Biological Assay; D001703:Biophysics / Q000379:methods*; D002696:Chlamydomonas / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008863:Microspheres; D008870:Microtubules / Q000378:metabolism; D036103:Nanotechnology; D052898:Optical Tweezers; D021381:Protein Transport; D034421:Sus scrofa
D005456:Fluorescent Dyes; D056444:Axonemal Dyneins