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来自咝蝰属蝰亚科蛇类毒液中的腺苷:使用液相色谱/质谱联用和毛细管电泳/质谱联用进行鉴定和定量分析

Adenosine in the venoms from viperinae snakes of the genus Bitis: identification and quantitation using LC/MS and CE/MS.

作者信息

Graham Robert Leslie James, McClean Stephen, O'Kane Edmund J, Theakston David, Shaw Chris

机构信息

School of Biomedical Sciences, University of Ulster, Coleraine, UK.

出版信息

Biochem Biophys Res Commun. 2005 Jul 22;333(1):88-94. doi: 10.1016/j.bbrc.2005.05.077.

Abstract

Snake venoms are rich sources of toxic proteins and small molecules. This study was directed at molecules of molecular mass below 1 kDa. Thirty different venoms, of either neurotoxic or haemorrhagic type, were fractionated using size-exclusion chromatography. Only venoms of the Puff adder (Bitis arietans), Gaboon viper (Bitis gabonica), and Rhinoceros viper (Bitis nasicornis) exhibited large absorbance peaks at lambda(280 nm) in the total volume range of the chromatographic column indicating the presence of abundant low molecular mass material. Analysis of fractions containing this material using both HPLC and capillary electrophoresis interfaced with electrospray ion-trap mass spectrometry unequivocally established that the bioactive nucleoside, adenosine, was the major component. The concentrations of adenosine found (Puff adder--97.7 x 10(-6) mol L(-1); Gaboon viper--28.0 x 10(-6) mol L(-1); and Rhinoceros viper-56.8 x 10(-6) mol L(-1)) were above those required to activate all known sub-types of adenosine receptors. Adenosine may thus act at the site of envenomation causing local vasodilatation and may play a role in the subsequent systemic hypotension observed.

摘要

蛇毒是有毒蛋白质和小分子的丰富来源。本研究针对分子量低于1 kDa的分子。使用尺寸排阻色谱法对30种不同的神经毒性或出血性蛇毒进行了分级分离。只有鼓腹咝蝰(Bitis arietans)、加蓬咝蝰(Bitis gabonica)和犀咝蝰(Bitis nasicornis)的蛇毒在色谱柱的总体积范围内于λ(280 nm)处出现大的吸光度峰,表明存在大量低分子量物质。使用高效液相色谱法和与电喷雾离子阱质谱联用的毛细管电泳法对含有该物质的级分进行分析,明确确定生物活性核苷腺苷是主要成分。所发现的腺苷浓度(鼓腹咝蝰——97.7×10⁻⁶ mol L⁻¹;加蓬咝蝰——28.0×10⁻⁶ mol L⁻¹;犀咝蝰——56.8×10⁻⁶ mol L⁻¹)高于激活所有已知腺苷受体亚型所需的浓度。因此,腺苷可能在蛇咬中毒部位起作用,导致局部血管扩张,并可能在随后观察到的全身性低血压中起作用。

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