Calmodulin interaction with cytoplasmic and flagellar dynein: calcium-dependent binding and stimulation of adenosinetriphosphatase activity.
Hisanaga and Sakai [Hisanaga, S., & Sakai, H. (1983) J. Biochem. (Tokyo) 93, 87-98] demonstrated that cytoplasmic dynein could be purified, in part, by chromatography on a calmodulin-Sepharose 4B affinity column and that the adenosinetriphosphatase (ATPase) activity of the enzyme was stimulated by Ca2+-calmodulin. In the present study, we have investigated, in detail, the interaction of cytoplasmic and flagellar dynein from the sea urchin Hemicentrotus pulcherrimus with calmodulin (CaM) isolated from porcine brain or sea urchin egg. The dynein Mg2+-ATPase activity is stimulated 3-8-fold by calmodulin from either source. The stimulation is dependent on calcium ions and is inhibited by trifluoroperazine. CaM stimulation is sensitive to physiologically regulatory calcium ion concentrations around 1 microM. Activation is also sensitive to pH and occurs maximally at physiological pH near 7.0. Calmodulin binds directly to cytoplasmic dynein as judged by cosedimentation in a sucrose density gradient. The binding and enzymatic stimulation occur at calmodulin:dynein ratios of 150:1 to 300:1, which are consistent with estimates of in vivo ratios. Cytoplasmic and flagellar dynein ATPase activities are also stimulated by Triton X-100, a nonionic detergent, and by limited protolysis with trypsin. Both of these treatments abolish further activation by calmodulin. The possibility of a trypsin-labile, CaM binding subunit of the enzyme is discussed. In addition, since both CaM and dynein are localized in the mitotic apparatus, we suggest that CaM may regulate possible mitotic dynein activity.
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000818:Animals; D001921:Brain; D017301:Ca(2+) Mg(2+)-ATPase; D002118:Calcium / Q000494:pharmacology*; D002147:Calmodulin / Q000378:metabolism*; D003902:Detergents / Q000494:pharmacology; D004398:Dyneins / Q000378:metabolism*; D005260:Female; D005407:Flagella / Q000201:enzymology*; D007700:Kinetics; D017830:Octoxynol; D010063:Ovum / Q000201:enzymology; D011092:Polyethylene Glycols / Q000494:pharmacology; D011485:Protein Binding; D012617:Sea Urchins; D013552:Swine; D014357:Trypsin
D002147:Calmodulin; D003902:Detergents; D011092:Polyethylene Glycols; D017830:Octoxynol; D014357:Trypsin; D000251:Adenosine Triphosphatases; D017301:Ca(2+) Mg(2+)-ATPase; D004398:Dyneins; D002118:Calcium