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不同盐类和去污剂对仙女虫(Fridericia heliota)荧光素 - 荧光素酶发光的影响。

Effect of different salts and detergents on luciferin-luciferase luminescence of the enchytraeid Fridericia heliota.

作者信息

Rodionova N S, Petushkov V N

机构信息

Institute of Biophysics, Siberian Division, Russian Academy of Sciences, Akademgorodok, Krasnoyarsk 660036, Russia.

出版信息

J Photochem Photobiol B. 2006 May 1;83(2):123-8. doi: 10.1016/j.jphotobiol.2005.12.014. Epub 2006 Feb 7.

Abstract

The study addresses the effect produced by different inorganic salts and detergents (SDS, Triton X-100, the Tween series) on the ATP-dependent bioluminescent reaction catalyzed by the luciferase of the new earthworm species Fridericia heliota (Annelida: Clitellata: Oligochaeta: Enchytraeidae). It has been shown that the effect of divalent metal salts on luminescence is determined by the action of cations. Three of them - Mg(2+), Mn(2+) and Ca(2+) - can stimulate luciferase activity at concentrations varying within a wide range, and Mn(2+) can act as a 100%-effective substitute for Mg(2+) in F. heliota luminescence reaction in vitro. The inhibitory effect of monovalent metal salts on luminescence is largely determined by the action of the anion part of the molecule. The effectiveness of the inhibitory effect of anions increases in the following order: Cl(-)<CO(3)(-2) approximately SO(3)(-2) approximately Br(-)<SO(4)(-2) approximately PO(4)(-3)<NO(3)(-)<I(-)<<Cr(2)O(7)(-2)<<Fe(CN)(6)(-3). Of the sodium salts, dodecyl sulfate, which is an anionic detergent, produces the strongest inhibitory effect on luciferase. On the contrary, nonionic detergents produce a stimulatory effect on the F. heliota luciferase. The action of the most effective of them - Triton X-100 - is determined by its ability to reduce the actual concentration of lipid inhibitors in the reaction mixture.

摘要

本研究探讨了不同无机盐和洗涤剂(十二烷基硫酸钠、曲拉通X-100、吐温系列)对新蚯蚓物种弗里德里西亚日光蚓(环节动物门:寡毛纲:线蚓科)荧光素酶催化的ATP依赖性生物发光反应的影响。研究表明,二价金属盐对发光的影响取决于阳离子的作用。其中三种——镁离子、锰离子和钙离子——在很宽的浓度范围内都能刺激荧光素酶活性,并且在体外弗里德里西亚日光蚓发光反应中,锰离子可以作为镁离子的100%有效替代物。一价金属盐对发光的抑制作用在很大程度上取决于分子阴离子部分的作用。阴离子抑制作用的有效性按以下顺序增加:氯离子<碳酸根离子≈亚硫酸根离子≈溴离子<硫酸根离子≈磷酸根离子<硝酸根离子<碘离子<重铬酸根离子<铁氰根离子。在钠盐中,作为阴离子洗涤剂的十二烷基硫酸钠对荧光素酶产生最强的抑制作用。相反,非离子洗涤剂对弗里德里西亚日光蚓荧光素酶产生刺激作用。其中最有效的曲拉通X-100的作用取决于其降低反应混合物中脂质抑制剂实际浓度的能力。

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