TY - JOUR
T1 - Three-headed outer arm dynein from Chlamydonwnas that can functionally combine with outer-arm-missing axonemes
AU - Takada, Saeko
AU - Sakakibara, Hitoshi
AU - Kamiya, Ritsu
PY - 1992/6
Y1 - 1992/6
N2 - A procedure was developed for Isolating Chlamydonzonas outer-arm dynein that can functionally combine with the axoneme of an outer-arm-missing mutant, oda1. Previous studies showed that the outer-arm dyneln of this organism, containing three heavy chains (α, β, γ), dissociates upon extraction with a high-salt-concentration buffer solution into an 18-S particle containing the α and β heavy chains and a 12-S particle containing the γ heavy chain. It was found, however, that the three heavy chains did not dissociate if the high-salt extract was centrifuged in the presence of Mg2+ the three chains constituted a single species (23-S dynein) sedimenting at about 23 S and displayed a three-headed bouquet configuration in electron micrographs. Furthermore, the 23-S dyneln had the activity to bind to the axonemes of oda1 and Increase the reactivated motility of detergent-extracted cell models; Its addition increased the beat frequency from 28Hz to 53 Hz, a frequency comparable to that of wild-type axoneme. The 18-S and 12-S dyneins, on the other hand, were unable to Increase the motility of oda1 axonemes even when added together. The new protocol thus enables purification of outer-arm dynein that retains Its functional activity. It will provide a useful experimental system with which to study the mechanism of outer-arm function.
AB - A procedure was developed for Isolating Chlamydonzonas outer-arm dynein that can functionally combine with the axoneme of an outer-arm-missing mutant, oda1. Previous studies showed that the outer-arm dyneln of this organism, containing three heavy chains (α, β, γ), dissociates upon extraction with a high-salt-concentration buffer solution into an 18-S particle containing the α and β heavy chains and a 12-S particle containing the γ heavy chain. It was found, however, that the three heavy chains did not dissociate if the high-salt extract was centrifuged in the presence of Mg2+ the three chains constituted a single species (23-S dynein) sedimenting at about 23 S and displayed a three-headed bouquet configuration in electron micrographs. Furthermore, the 23-S dyneln had the activity to bind to the axonemes of oda1 and Increase the reactivated motility of detergent-extracted cell models; Its addition increased the beat frequency from 28Hz to 53 Hz, a frequency comparable to that of wild-type axoneme. The 18-S and 12-S dyneins, on the other hand, were unable to Increase the motility of oda1 axonemes even when added together. The new protocol thus enables purification of outer-arm dynein that retains Its functional activity. It will provide a useful experimental system with which to study the mechanism of outer-arm function.
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U2 - 10.1093/oxfordjournals.jbchem.a123832
DO - 10.1093/oxfordjournals.jbchem.a123832
M3 - Article
C2 - 1386849
AN - SCOPUS:0026681097
SN - 0021-924X
VL - 111
SP - 758
EP - 762
JO - Journal of Biochemistry
JF - Journal of Biochemistry
IS - 6
ER -