材料期刊网

家福捕鸟蛛（ Selenocosmia jiafu）是一种生活在中国广西、云南等边远山区、中等个体、产毒量较大和毒性较强的蜘蛛新种。为了对家福捕鸟蛛粗毒成分进行初步探索，采用反相高效液相色谱、基质辅助激光解吸离子化飞行时间质谱和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳方法对粗毒多肽和蛋白质的多样性进行了分析。结果表明：家福捕鸟蛛粗毒经色谱分离后得到40多个色谱峰，经质谱鉴定得到238个多肽，且多肽的相对分子质量呈现出双峰分布，其中62.5％的多肽的相对分子质量分布在3000~4500之间，33.2％的多肽的相对分子质量分布在1000~3000之间。这种相对分子质量的分布模式不同于其他已经报道的蜘蛛粗毒中多肽的分布模式。电泳分析结果表明：除了相对分子质量在10000以下的多肽分子，粗毒在50、72和90 kD附近有3条明显的条带，粗毒电泳条带经液相色谱-电喷雾四极杆飞行时间质谱鉴定，主要是一些血蓝蛋白、钾离子通道蛋白、钙蛋白酶等。说明家福捕鸟蛛粗毒中多肽和蛋白质种类丰富。

Selenocosmia jiafu( S. jiafu)is recently identified as a new species of spider in P. R. China. These medium bodied venomous spiders are distributed mainly in the hilly areas of southwest of China,mostly at Yunnan and Guangxi Provinces. In order to understand the composition of the S. jiafu venom,we performed a preliminary analysis of this venom using reversed-phase high performance liquid chromatography ( RP-HPLC ), matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry( MALDI-TOF-MS)and sodium dodecyl sulfate-polyacrylamide gel electrophoresis( SDS-PAGE). The S. jiafu venom was separated by RP-HPLC in an analytical C18 column(phenomenex 100 ?,250 mm×4. 6 mm,5 μm)equilibra-ted with solution A( distilled water with 0. 1% trifluoroacetic acid),using a gradient from 0%to 50% of solution B( acetonitrile with 0. 1% trifluoroacetic acid)over 50 min with a flow rate of 1 mL/min. The isolated venom proteins were treated with in-gel digestion separated by SDS-PAGE and then identified by liquid chromatography-electrospray ionization quadrupole-time of flight mass spectrometry( LC-ESI-QTOF-MS)techniques. The results show that more than 40 fractions eluted were monitored at 215 nm in the RP-HPLC chromatogram of the venom of the spider S. jiafu. Most of the fractions were eluted with retention times of 5-15 min and 25-40 min,corresponding to 5% -15% and 25% -40% acetonitrile,respectively. The venom contains 238 peptides that follow a bimodal distribution,with about 62. 5% of the peptides having a rela-tive molecular mass of 3 000-4 500 and about 33. 2% of the peptides having a relative molecular mass of 1 000-3 000. This distribution model is rather different from those of peptides from oth-er tarantula spider venoms analyzed. To explore the relative molecular mass distribution of the venom proteins,the venom was analyzed by SDS-PAGE using standard protocols. Except for peptides with relative molecular mass lower than 10 000, the SDS-PAGE electrophoresis revealed three more obvious bands around 50,72 and 90 kD respectively. Further MS analysis indicated that there are mainly hemocyanin,potassium ion channel protein,calcium protease and so on. Altogether,this study not only indicated there are many peptides and proteins in the S. jiafu venom,but also provided a basis for further case-by-case investigation of peptide tox-ins from this venom.