Stimulatory effects of arachidonic acid on myosin ATPase activity and contraction of smooth muscle via myosin motor domain

Takeshi Katayama, Masaru Watanabe, Hideyuki Tanaka, Mizuki Hino, Takuya Miyakawa, Takashi Ohki, Li Hong Ye, Ce Xie, Shinji Yoshiyama, Akio Nakamura, Ryoki Ishikawa, Masaru Tanokura, Kazuhiro Oiwa, Kazuhiro Kohama

    Research output: Contribution to journalArticle

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    Abstract

    We have been searching for a mechanism to induce smooth muscle contraction that is not associated with phosphorylation of the regulatory light chain (RLC) of smooth muscle myosin (Nakamura A, Xie C, Zhang Y, Gao Y, Wang HH, Ye LH, Kishi H, Okagaki T, Yoshiyama S, Hayakawa K, Ishikawa R, Kohama K. Biochem Biophys Res Commun 369: 135-143, 2008). In this article, we report that arachidonic acid (AA) stimulates ATPase activity of unphosphorylated smooth muscle myosin with maximal stimulation (R max) of 6.84 ± 0.51 relative to stimulation by the vehicle and with a half-maximal effective concentration (EC 50) of 50.3 ± 4.2 μM. In the presence of actin, R max was 1.72 ± 0.08 and EC 50 was 26.3 ± 2.3 μM. Our experiments with eicosanoids consisting of the AA cascade suggested that they neither stimulated nor inhibited the activity. Under conditions that did not allow RLC to be phosphorylated, AA stimulated contraction of smooth muscle tissue with an R max of 1.45 ± 0.07 and an EC 50 of 27.0 ± 4.4 μM. In addition to the ATPase activities of the myosin, AA stimulated those of heavy meromyosin, subfragment 1 (S1), S1 from which the RLC was removed, and a recombinant heavy chain consisting of the myosin head. The stimulatory effects of AA on these preparations were about twofold. The site of AA action was indicated to be the step-releasing inorganic phosphate (P i) from the reaction intermediate of the myosin-ADP-P i complex. The enhancement of Pi release by AA was supported by computer simulation indicating that AA docked in the actin-binding cleft of the myosin motor domain. The stimulatory effect of AA was detectable with both unphosphorylated myosin and the myosin in which RLC was fully phosphorylated. The AA effect on both myosin forms was suggested to cause excess contraction such as vasospasm.

    Original languageEnglish
    JournalAmerican Journal of Physiology - Heart and Circulatory Physiology
    Volume298
    Issue number2
    DOIs
    Publication statusPublished - 2010

    Fingerprint

    Smooth Muscle Myosins
    Myosins
    Arachidonic Acid
    Light
    Smooth Muscle
    Actins
    Myosin Subfragments
    Myosin Heavy Chains
    Eicosanoids
    Muscle Contraction
    Computer Simulation
    Adenosine Diphosphate
    Adenosine Triphosphatases
    Phosphates
    Phosphorylation

    Keywords

    • Skinned fiber

    ASJC Scopus subject areas

    • Physiology
    • Physiology (medical)
    • Cardiology and Cardiovascular Medicine

    Cite this

    Stimulatory effects of arachidonic acid on myosin ATPase activity and contraction of smooth muscle via myosin motor domain. / Katayama, Takeshi; Watanabe, Masaru; Tanaka, Hideyuki; Hino, Mizuki; Miyakawa, Takuya; Ohki, Takashi; Ye, Li Hong; Xie, Ce; Yoshiyama, Shinji; Nakamura, Akio; Ishikawa, Ryoki; Tanokura, Masaru; Oiwa, Kazuhiro; Kohama, Kazuhiro.

    In: American Journal of Physiology - Heart and Circulatory Physiology, Vol. 298, No. 2, 2010.

    Research output: Contribution to journalArticle

    Katayama, T, Watanabe, M, Tanaka, H, Hino, M, Miyakawa, T, Ohki, T, Ye, LH, Xie, C, Yoshiyama, S, Nakamura, A, Ishikawa, R, Tanokura, M, Oiwa, K & Kohama, K 2010, 'Stimulatory effects of arachidonic acid on myosin ATPase activity and contraction of smooth muscle via myosin motor domain', American Journal of Physiology - Heart and Circulatory Physiology, vol. 298, no. 2. https://doi.org/10.1152/ajpheart.00577.2009
    Katayama, Takeshi ; Watanabe, Masaru ; Tanaka, Hideyuki ; Hino, Mizuki ; Miyakawa, Takuya ; Ohki, Takashi ; Ye, Li Hong ; Xie, Ce ; Yoshiyama, Shinji ; Nakamura, Akio ; Ishikawa, Ryoki ; Tanokura, Masaru ; Oiwa, Kazuhiro ; Kohama, Kazuhiro. / Stimulatory effects of arachidonic acid on myosin ATPase activity and contraction of smooth muscle via myosin motor domain. In: American Journal of Physiology - Heart and Circulatory Physiology. 2010 ; Vol. 298, No. 2.
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    AU - Hino, Mizuki

    AU - Miyakawa, Takuya

    AU - Ohki, Takashi

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    AU - Yoshiyama, Shinji

    AU - Nakamura, Akio

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