Equilibrium and transition between single- and double-headed binding of kinesin as revealed by single-molecule mechanics

Kenji Kawaguchi, Sotaro Uemura, Shin'Ichi Ishiwata

    Research output: Contribution to journalArticle

    38 Citations (Scopus)

    Abstract

    Kinesin is a processive motor protein that "walks" on a microtubule toward its plus end. We reported previously that the distribution of unbinding force and elastic modulus for a single kinesin-microtubule complex was either unimodal or bimodal depending on the nucleotide states of the kinesin heads, hence showing that the kinesin may bind the microtubule either with one head or with both heads at once. Here, we found that the shape of the unbinding-force distribution depends both on the loading rate and on the manner of loading not only in the presence of AMP-PNP but also in the absence of nucleotides. Irrespective of the nucleotide state and the loading conditions examined here, the unbinding force obtained by loading directed toward the minus end of microtubule was 45% greater than that for plus end-directed loading. These results could be explained by a model in which equilibrium exists between single- and double-headed binding and the load (F) dependence of lifetime, τ(F), of each binding is expressed by τ(F) = τ(0)exp(-Fd/kBT), where τ(0) is the lifetime without external load and d a characteristic distance, both of which depend on single- or double-headed binding, kB, the Boltzmann constant and T, the absolute temperature. The model analysis showed that the forward and backward rates of transition from single- to doubleheaded binding are 2 and 0.2/s for the AMP-PNP state, and 70 and 7/s for the nucleotide-free state. Moreover, in the presence of AMP-PNP, we detected the moment of transition from single- to double-headed binding through an abrupt increase in the elastic modulus and estimated the transition rate to be ∼1/s, which is consistent with the model analysis.

    Original languageEnglish
    Pages (from-to)1103-1113
    Number of pages11
    JournalBiophysical Journal
    Volume84
    Issue number2 I
    Publication statusPublished - 2003 Feb 1

    Fingerprint

    Kinesin
    Adenylyl Imidodiphosphate
    Mechanics
    Microtubules
    Nucleotides
    Elastic Modulus
    Head
    Theophylline
    Temperature
    Proteins

    ASJC Scopus subject areas

    • Biophysics

    Cite this

    Equilibrium and transition between single- and double-headed binding of kinesin as revealed by single-molecule mechanics. / Kawaguchi, Kenji; Uemura, Sotaro; Ishiwata, Shin'Ichi.

    In: Biophysical Journal, Vol. 84, No. 2 I, 01.02.2003, p. 1103-1113.

    Research output: Contribution to journalArticle

    Kawaguchi, Kenji ; Uemura, Sotaro ; Ishiwata, Shin'Ichi. / Equilibrium and transition between single- and double-headed binding of kinesin as revealed by single-molecule mechanics. In: Biophysical Journal. 2003 ; Vol. 84, No. 2 I. pp. 1103-1113.
    @article{53357336eebc4c57af84f8a03da164dd,
    title = "Equilibrium and transition between single- and double-headed binding of kinesin as revealed by single-molecule mechanics",
    abstract = "Kinesin is a processive motor protein that {"}walks{"} on a microtubule toward its plus end. We reported previously that the distribution of unbinding force and elastic modulus for a single kinesin-microtubule complex was either unimodal or bimodal depending on the nucleotide states of the kinesin heads, hence showing that the kinesin may bind the microtubule either with one head or with both heads at once. Here, we found that the shape of the unbinding-force distribution depends both on the loading rate and on the manner of loading not only in the presence of AMP-PNP but also in the absence of nucleotides. Irrespective of the nucleotide state and the loading conditions examined here, the unbinding force obtained by loading directed toward the minus end of microtubule was 45{\%} greater than that for plus end-directed loading. These results could be explained by a model in which equilibrium exists between single- and double-headed binding and the load (F) dependence of lifetime, τ(F), of each binding is expressed by τ(F) = τ(0)exp(-Fd/kBT), where τ(0) is the lifetime without external load and d a characteristic distance, both of which depend on single- or double-headed binding, kB, the Boltzmann constant and T, the absolute temperature. The model analysis showed that the forward and backward rates of transition from single- to doubleheaded binding are 2 and 0.2/s for the AMP-PNP state, and 70 and 7/s for the nucleotide-free state. Moreover, in the presence of AMP-PNP, we detected the moment of transition from single- to double-headed binding through an abrupt increase in the elastic modulus and estimated the transition rate to be ∼1/s, which is consistent with the model analysis.",
    author = "Kenji Kawaguchi and Sotaro Uemura and Shin'Ichi Ishiwata",
    year = "2003",
    month = "2",
    day = "1",
    language = "English",
    volume = "84",
    pages = "1103--1113",
    journal = "Biophysical Journal",
    issn = "0006-3495",
    publisher = "Biophysical Society",
    number = "2 I",

    }

    TY - JOUR

    T1 - Equilibrium and transition between single- and double-headed binding of kinesin as revealed by single-molecule mechanics

    AU - Kawaguchi, Kenji

    AU - Uemura, Sotaro

    AU - Ishiwata, Shin'Ichi

    PY - 2003/2/1

    Y1 - 2003/2/1

    N2 - Kinesin is a processive motor protein that "walks" on a microtubule toward its plus end. We reported previously that the distribution of unbinding force and elastic modulus for a single kinesin-microtubule complex was either unimodal or bimodal depending on the nucleotide states of the kinesin heads, hence showing that the kinesin may bind the microtubule either with one head or with both heads at once. Here, we found that the shape of the unbinding-force distribution depends both on the loading rate and on the manner of loading not only in the presence of AMP-PNP but also in the absence of nucleotides. Irrespective of the nucleotide state and the loading conditions examined here, the unbinding force obtained by loading directed toward the minus end of microtubule was 45% greater than that for plus end-directed loading. These results could be explained by a model in which equilibrium exists between single- and double-headed binding and the load (F) dependence of lifetime, τ(F), of each binding is expressed by τ(F) = τ(0)exp(-Fd/kBT), where τ(0) is the lifetime without external load and d a characteristic distance, both of which depend on single- or double-headed binding, kB, the Boltzmann constant and T, the absolute temperature. The model analysis showed that the forward and backward rates of transition from single- to doubleheaded binding are 2 and 0.2/s for the AMP-PNP state, and 70 and 7/s for the nucleotide-free state. Moreover, in the presence of AMP-PNP, we detected the moment of transition from single- to double-headed binding through an abrupt increase in the elastic modulus and estimated the transition rate to be ∼1/s, which is consistent with the model analysis.

    AB - Kinesin is a processive motor protein that "walks" on a microtubule toward its plus end. We reported previously that the distribution of unbinding force and elastic modulus for a single kinesin-microtubule complex was either unimodal or bimodal depending on the nucleotide states of the kinesin heads, hence showing that the kinesin may bind the microtubule either with one head or with both heads at once. Here, we found that the shape of the unbinding-force distribution depends both on the loading rate and on the manner of loading not only in the presence of AMP-PNP but also in the absence of nucleotides. Irrespective of the nucleotide state and the loading conditions examined here, the unbinding force obtained by loading directed toward the minus end of microtubule was 45% greater than that for plus end-directed loading. These results could be explained by a model in which equilibrium exists between single- and double-headed binding and the load (F) dependence of lifetime, τ(F), of each binding is expressed by τ(F) = τ(0)exp(-Fd/kBT), where τ(0) is the lifetime without external load and d a characteristic distance, both of which depend on single- or double-headed binding, kB, the Boltzmann constant and T, the absolute temperature. The model analysis showed that the forward and backward rates of transition from single- to doubleheaded binding are 2 and 0.2/s for the AMP-PNP state, and 70 and 7/s for the nucleotide-free state. Moreover, in the presence of AMP-PNP, we detected the moment of transition from single- to double-headed binding through an abrupt increase in the elastic modulus and estimated the transition rate to be ∼1/s, which is consistent with the model analysis.

    UR - http://www.scopus.com/inward/record.url?scp=0037306229&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=0037306229&partnerID=8YFLogxK

    M3 - Article

    C2 - 12547791

    AN - SCOPUS:0037306229

    VL - 84

    SP - 1103

    EP - 1113

    JO - Biophysical Journal

    JF - Biophysical Journal

    SN - 0006-3495

    IS - 2 I

    ER -