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过钼酸盐的特性及其对细胞酪氨酸磷酸化、间隙连接细胞间通讯以及间隙连接蛋白连接蛋白43磷酸化状态的影响。

A characterization of permolybdate and its effect on cellular tyrosine phosphorylation, gap junctional intercellular communication and phosphorylation status of the gap junction protein, connexin43.

作者信息

Mikalsen S O, Kaalhus O

机构信息

Department for Environmental and Occupational Cancer, Institute for Cancer Research, The Norwegian Radium Hospital, Oslo.

出版信息

Biochim Biophys Acta. 1997 Apr 24;1356(2):207-20. doi: 10.1016/s0167-4889(96)00163-2.

Abstract

Biological and analytical characterizations of permolybdate (a mixture of H2O2 and molybdate) were done. Molybdate (10 mM) and molybdenum(V) chloride (3 mM) did not affect gap junctional intercellular communication (GJIC), phosphorylation status of connexin43 (Cx43) or cellular tyrosine phosphorylation in early passage hamster embryonic cells (mainly fibroblast-like). High concentrations of H2O2 (3-10 mM) affected some of the parameters. Acidified permolybdate was clearly more stable than the unadjusted permolybdate. The maximum biological potency of acidified permolybdate was found at a molar ratio of 2:1 (H2O2:molybdate). The mixtures of molybdenum(V) chloride and H2O2 gave a maximum effect at 4:1 molar ratio (H2O2:molybdenum(V)). This can be explained by decomposition of H2O2 and by the generation of less biologically active compounds. Spectrophotometric analyses of the mixtures corroborated the biological results. The Mo(V) electron spin resonance spectrum disappeared upon addition of H2O2 to Mo(V) solutions, and no spectrum appeared when H2O2 was mixed with Mo(VI). Thus, permolybdate is probably diperoxomolybdate, a Mo(VI) compound. Regardless of the parent metal salt, the H2O2/metal salt mixtures showed concentration-dependent biphasic responses with an initial decrease in GJIC followed by an increase. A dissociation between alteration in Cx43 phosphorylation status and GJIC was obtained under certain conditions. The biological activities of permolybdate were only partially mimicked by phenylarsine oxide, an alternative protein tyrosine phosphatase inhibitor.

摘要

对过钼酸盐(过氧化氢和钼酸盐的混合物)进行了生物学和分析表征。钼酸盐(10 mM)和氯化钼(V)(3 mM)不影响早期传代仓鼠胚胎细胞(主要是成纤维样细胞)中的间隙连接细胞间通讯(GJIC)、连接蛋白43(Cx43)的磷酸化状态或细胞酪氨酸磷酸化。高浓度的过氧化氢(3 - 10 mM)会影响一些参数。酸化的过钼酸盐明显比未调节的过钼酸盐更稳定。酸化过钼酸盐的最大生物活性在摩尔比为2:1(过氧化氢:钼酸盐)时被发现。氯化钼(V)和过氧化氢的混合物在摩尔比为4:1(过氧化氢:钼(V))时产生最大效果。这可以通过过氧化氢的分解以及生成生物活性较低的化合物来解释。混合物的分光光度分析证实了生物学结果。向钼(V)溶液中加入过氧化氢后,钼(V)电子自旋共振谱消失,而过氧化氢与钼(VI)混合时没有出现谱线。因此,过钼酸盐可能是双氧钼酸盐,一种钼(VI)化合物。无论母体金属盐如何,过氧化氢/金属盐混合物都表现出浓度依赖性的双相反应,最初GJIC下降,随后上升。在某些条件下,Cx43磷酸化状态的改变与GJIC之间出现了解离。过钼酸盐的生物活性仅部分被氧化苯砷(一种替代的蛋白酪氨酸磷酸酶抑制剂)模拟。

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