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Literature summary for 5.6.1.2 extracted from

  • Imamula, K.; Kon, T.; Ohkura, R.; Sutoh, K.
    The coordination of cyclic microtubule association/dissociation and tail swing of cytoplasmic dynein (2007), Proc. Natl. Acad. Sci. USA, 104, 16134-16139.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression of C-terminal 380-kDa motor domain of cytoplasmic dynein heavy chain fused with His6-FLAG-GFP at its N-terminus and BFP at the interior of the AAA2 module, and fused with a His6-FLAG-biotinylation sequence at its N terminus Dictyostelium discoideum

Protein Variants

Protein Variants Comment Organism
E2022Q mutation in the Walker B motif of the AAA1 module. In the presence of ATP, the mutant is trapped in a single state, D*-ATP. It binds to microtubules with high affintiy, similar to wild-type Dictyostelium discoideum
K1975T Walker A mutation in the AAA1 module. Mutant tightly binds to microtubules in the presence and absence of ATP. The inhibition of ATP binding to the AAA1 ATPase site by the mutation completely blocks the ATP-induced dissociation of the complex with the microtubules Dictyostelium discoideum
K2675T Walker A mutation in the AAA1 module. Mutant tightly binds to microtubules in the presence and absence of ATP. Inhibition of ATP binding to the AAA3 ATPase site by the K2675T mutationdoes not block the ATP-induced dissociation of the complex with the microtubules Dictyostelium discoideum

Organism

Organism UniProt Comment Textmining
Dictyostelium discoideum
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Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information at most of the intermediate states in the ATPase cycle, two mechanical processes proceed simultaneously: the motor domain in the poststroke state binds strongly to the microtubule with a Kd of 0.2 microM, whereas most of the motor domains in the prestroke state bind weakly to the microtubule with a Kd of >10 microM. The timings of the microtubule affinity change and tail swing are staggered at the recovery stroke step in which the tail swings from the poststroke to the prestroke position Dictyostelium discoideum ?
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