Interaction between src family kinases and rho-kinase in agonist-induced Ca2+-sensitization of rat pulmonary artery.
Interaction between src family kinases and rho-kinase in agonist-induced Ca2+-sensitization of rat pulmonary artery.
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DOI：
10.1093/cvr/cvm073
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发表时间：
2008-02-01
影响因子：
10.8
通讯作者：
Aaronson PI
中科院分区：
文献类型：
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作者：
Knock GA;Shaifta Y;Snetkov VA;Vowles B;Drndarski S;Ward JP;Aaronson PI
我们研究了src家族激酶（srcFK）在激动剂介导的肺动脉Ca2+敏化中的作用，以及这是否涉及与rho/rho激酶途径的相互作用。获得大鼠肺内动脉（IPAs）和培养肺动脉平滑肌细胞（PASMC）。在mRNA和蛋白水平上检测srcFK的表达。前列腺素F2α (PGF2α)在α-毒素渗透IPAs中诱导Ca2+敏化。我们还检测了肌球蛋白磷酸酶（MYPT-1）和肌球蛋白轻链-20 （MLC20）调控亚基的磷酸化和rhok易位对PGF2α的响应。9种srcFK mRNA水平表达，包括src、fyn和yes， PGF2α增强了3种srcFK蛋白tyr1 -416位点的磷酸化。在α-毒素渗透IPAs中，PGF2α使Ca2+诱导的收缩（pCa 6.9）增强约3倍。这种增强被srcFK阻断剂SU6656和PP2以及rho激酶抑制剂Y27632所抑制。Y27632也能抑制潜在的pCa 6.9收缩，但SU6656和PP2没有。PGF2α增强了MYPT-1在thr-697和thr-855位点的磷酸化，以及MLC20在ser-19位点的磷酸化。这种增强，而不是潜在的基础磷酸化，被SU6656抑制。Y27632抑制基础磷酸化和pgf2 α介导的磷酸化。SU6656和Y27632对MYPT-1和MLC20磷酸化的影响并不是相加的。PGF2α触发PASMC中rho激酶的易位，而SU6656可以抑制这种易位。srcFK在大鼠肺动脉中被PGF2α激活，并可能通过rhok易位和MYPT-1磷酸化参与Ca2+敏化和收缩。
We investigated the role of src family kinases (srcFK) in agonist-mediated Ca2+-sensitization in pulmonary artery and whether this involves interaction with the rho/rho-kinase pathway. Intra-pulmonary arteries (IPAs) and cultured pulmonary artery smooth muscle cells (PASMC) were obtained from rat. Expression of srcFK was determined at the mRNA and protein levels. Ca2+-sensitization was induced by prostaglandin F2α (PGF2α) in α-toxin-permeabilized IPAs. Phosphorylation of the regulatory subunit of myosin phosphatase (MYPT-1) and of myosin light-chain-20 (MLC20) and translocation of rho-kinase in response to PGF2α were also determined. Nine srcFK were expressed at the mRNA level, including src, fyn, and yes, and PGF2α enhanced phosphorylation of three srcFK proteins at tyr-416. In α-toxin-permeabilized IPAs, PGF2α enhanced the Ca2+-induced contraction (pCa 6.9) approximately three-fold. This enhancement was inhibited by the srcFK blockers SU6656 and PP2 and by the rho-kinase inhibitor Y27632. Y27632, but not SU6656 or PP2, also inhibited the underlying pCa 6.9 contraction. PGF2α enhanced phosphorylation of MYPT-1 at thr-697 and thr-855 and of MLC20 at ser-19. This enhancement, but not the underlying basal phosphorylation, was inhibited by SU6656. Y27632 suppressed both basal and PGF2α-mediated phosphorylation. The effects of SU6656 and Y27632, on both contraction and MYPT-1 and MLC20 phosphorylation, were not additive. PGF2α triggered translocation of rho-kinase in PASMC, and this was inhibited by SU6656. srcFK are activated by PGF2α in the rat pulmonary artery and may contribute to Ca2+-sensitization and contraction via rho-kinase translocation and phosphorylation of MYPT-1.