Partial Purification and Characterization of Dynein Adenosine Triphosphatase from Bovine Sperm1

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Abstract
Outer dynein and polypeptides that possess Mg2+-adenosine triphosphatase (ATPase) activity have been extracted from the flagellar axonemes of demembranated bovine sperm. Electron microscopy of intact and salt-extracted sperm demonstrates a relatively selective removal of the outer dynein arms. The salt extract contains a specific ATPase activity of 55 nmoles inorganic phosphate (Pi)/min/mg protein. Sucrose density gradient centrifugation of this extract results in a 6-fold increase in specific sactivity of ATPase (333 nmole/Pi/min/mg protein), which sediments as a single 13S peak. Concomitant with the increase in specific activity, there is enrichment of three high molecular weight polypeptides (Mr > 300,000) characteristic of dynein heavy chains. ATPase activities in the initial extract and in the 113S peak are inhibited by concentrations of vanadate and erythro-9-[3-2-(hydroxynonyl)] adenine similar to those that inhibit ATPase activity in sea urchin sperm dynein. These findings indicate that outer arm dynein ATPase can be extracted and partially purified from bovine sperm.