Inner arm dyneins from flagella of Chlamydomonas reinhardtii.
Two dyneins have been isolated from axonemes of Chlamydomonas flagella by a three step procedure consisting of extraction in a high salt containing buffer, hydroxyapatite chromatography and sedimentation in sucrose gradient. A dynein with Mg+2- dependent ATPase activity 6.0 mumole Pi/min/mg, sedimenting at 12.5S was found associated with a polypeptide of molecular weight 310,000. A second dynein with specific activity of 3.7, sedimenting at 10-11S was found associated with a polypeptide of molecular weight 315,000. In their most purified forms, the two dyneins are complexed with nonstoichiometric amounts of four polypeptides ranging in molecular weight between 42,000 and 19,000. The 42,000 component has been identified previously as an actin-like protein. The high molecular weight subunits of both dyneins and two polypeptides of 28,000 and 19,000 molecular weight were found to be phosphorylated by in vivo pulse-labeling with 32P-phosphoric acid. All components of the 12.5S and 10-11S dynein complexes, with the exception of the 19,000 polypeptide, form a subset of polypeptides found to be deficient in pf-23, a chlamydomonas mutant, which is defective for inner arms.
D000251:Adenosine Triphosphatases / Q000032:analysis / Q000302:isolation & purification*; D002499:Centrifugation, Density Gradient; D002696:Chlamydomonas; D002850:Chromatography, Gel; D004398:Dyneins / Q000302:isolation & purification* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005407:Flagella / Q000032:analysis*; D010455:Peptides / Q000032:analysis; D010766:Phosphorylation
D010455:Peptides; D000251:Adenosine Triphosphatases; D004398:Dyneins