【作者】 赵丽萍；

【导师】 肖杭；

【作者基本信息】 南京医科大学 ， 卫生毒理学， 2005， 硕士

【摘要】 第一部分 东亚钳蝎毒素的分离纯化及性质鉴定 本实验室运用经典色谱法对东亚钳蝎(Buthus martensi Karsch, BmK)蝎毒素多肽进行了分离和纯化,最后得到了BmK2-2-5-5、BmK2-2-5-5-1以及BmK2-2-5-6、BmK2-2-5-6-1四个多肽单体,并且对所分离纯化的多肽的基本性质进行了初步鉴定。结果显示:(1)BmK2-2-5-5-1和BmK2-2-5-6-1对紫外扫描呈末端吸收,其吸收峰均在206nm左右;两者的分子量分别为7214.36 Da、7036.85 Da。(2)BmK2-2-5-5-1氨基酸分析的结果提示其残基个数的大致比例:Asp:Thr:Ser:Glu:Gly:Ala:Val:Met:Ile:Leu:Tyr:Phe:Lys:His:Arg≈4:1:8:4:10:4:4:2:2:2:2:2:2:1:2。(3) 运用Edman降解法测序,得到了BmK2-2-5-6的序列,为GRDAYIADSE NCTYT;其它三个单体因更多还原

【Abstract】 Part one: The purification, isolation and identification of Buthus martensi Karsch(BmK) neurotoxinsBmK2-2-5-5, BmK2-2-5-5-l and BmK2-2-5-6, BmK2-2-5-6-l were purified and isolated from scorpion crude venom by classical chromatographic method, and we studied the primary characterization and identification of these peptides. It showed: (1) Both of BmK2-2-5-5-l and BmK2-2-5-6-l revealed end absorption by UV scan, which the absorbency is the highest at 206nm or so. The MW of the twocomponents are 7214.36 Da and 7036.85 Da. (2) Acid amino analysis was used to determinate the relative ratio of the acid amino residues of BmK2-2-5-5-l: Asp: Thr: Ser: Glu: Gly: Ala: Val: Met: Iie: Leu: Tyr: Phe: Lys: His: Arg≈4: 1: 8: 4: 10: 4: 4: 2: 2: 2: 2: 2: 2: 1: 2. (3) The sequence of BmK2-2-5-6 is GRDAY IADSE NCTYT by Edman degradation; and those three peptides did not detect any signals because of N-terminal blocked.Part two: Effects of BmK2-2-5-5 and BmK2-2-5-5-l on Na~+currents and K~+currents in rat dorsal root ganglion neuronsThe effects of BmK2-2-5-5 and BmK2-2-5-5-l, polypeptides purified from Buthus martensi Karsch (BmK) venom, on Na~+ currents and K~+ currents in rat DRG neurons were investigated in this part using Whole-cell patch clamping technique. The results indicated as follows: (1) BmK2-2-5-5 does not exert significant effects on the sodium channel and the peak Na~+ currents at 50μg/ml; but at the concentration of 75μg/ml BmK2-2-5-5 showed significant increasing effects on the potassium channel in rat DRG neurons, while the outward potassium currents, the delayed rectifier potassium currents and the fast inactivating potassium currents were increased to 40.32 ± 10.13%(n=3), 33.22 ± 10.59%(n=3)更多还原