Purification and characterization of Salmo gairdneri outer arm dynein

Abstract

Dyneins are multimeric ATPases, which make up the inner and outer arms that bridge the outer doublet microtubules of eukaryotic cilia and flagella. They are responsible for the generation of sliding between outer doublets, which in turn is the basis for the formation and propagation of bending waves in both cilia and flagella. Outer arm dyneins are composed of two to three ATPases of Mr > 400,000, referred to as the α, β, and, where appropriate, γ heavy chains. Trout sperm is a new source of vertebrate dynein. Sperm can be repeatedly obtained in large quantities from the same trout (up to 4 × 10 spermatozoa per ejaculate), their axonemes can be readily isolated, and the dynein can be extracted efficiently and without significant proteolytic degradation. The advantages of trout sperm have permitted the detailed characterization of trout outer arm dynein to progress rapidly, so that it is now one of the best characterized of all dyneins. This chapter presents an overview of trout physiology and spermatogenesis for those not well acquainted with teleost physiology and anatomy and describes the methods for purification and characterization of Salmo gairdneri outer arm dynein.