What question did this study set out to answer?

To determine whether fluorescent 1,N6-etheno-ATP can serve as a nonperturbing tool to analyze the kinetics and mechanisms of steady-state subunit exchange in F-actin filaments.

September 1, 1981Proceedings of the National Academy of SciencesOpen Access

Exchange of 1,N6-etheno-ATP with actin-bound nucleotides as a tool for studying the steady-state exchange of subunits in F-actin solutions.

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Population

F-actin and G-actin solutions

Comparison

Addition of 1-N6-ethenoadenosine 5'-triphosphate vs Various conditions including phalloidin…

Design

Preclinical

Authors

YWY L WangAustrian Academy of Sciences DTD. Lansing TaylorUniversity of Pittsburgh

Discussion

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Implication

epsilon-ATP assay offers a nonperturbing probe for actin turnover; leaves open validation in cardiovascular cell and tissue models.

Key Points

To determine whether fluorescent 1,N6-etheno-ATP can serve as a nonperturbing tool to analyze the kinetics and mechanisms of steady-state subunit exchange in F-actin filaments.
Measured fluorescence intensity enhancement upon the binding of 1,N6-ethenoadenosine 5'-triphosphate (epsilon-ATP) to G-actin and steady-state F-actin solutions.
Evaluated exchange kinetics under mechanical shearing and pharmacological modulation with phalloidin or cytochalasin B (0.1 to 10 microM).
Characterized directional subunit assembly and disassembly using pulse-chase fluorescence assays in the presence and absence of millimolar magnesium ions.
Identified biphasic exchange kinetics comprising a fast phase and a slow phase, both of which were stimulated by shearing and inhibited by phalloidin.
Demonstrated via pulse-chase assays that actin subunits assemble and disassemble at the same filament end during the fast phase, but turnover occurs at opposite ends during the slow phase.
Observed that the slow phase requires millimolar magnesium ions and remains uninhibited by cytochalasin B at concentrations between 0.1 and 10 microM.

Structured PICO

Population

F-actin and G-actin solutions

Intervention

Addition of 1-N6-ethenoadenosine 5'-triphosphate (epsilon-ATP)

Comparator

Various conditions including phalloidin treatment, absence of millimolar magnesium ions, and cytochalasin B (0.1 to 10 microM)

Outcome

Kinetics of nucleotide exchange measured by fluorescence intensitysurrogate

The exchange of epsilon-ATP provides a nonperturbing tool to study the steady-state exchange of free actin subunits with subunits in actin filaments.

Cite This Study

Wang et al. (1981) studied this question.

synapsesocial.com/papers/6a91240dd2f2d69b3a7467dd https://doi.org/10.1073/pnas.78.9.5503

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Exchange of 1,N6-etheno-ATP with actin-bound nucleotides as a tool for studying the steady-state exchange of subunits in F-actin solutions.

Discussion

Member takes

Implication

Key Points

Structured PICO

Cite This Study

Exchange of 1,N6-etheno-ATP with actin-bound nucleotides as a tool for studying the steady-state exchange of subunits in F-actin solutions.

Discussion

Member takes

Implication

Key Points

Structured PICO

Cite This Study

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