MALDI-TOF质谱技术分析加州海兔口腔神经节中超微量Leu-Leu多肽酶

Analysis of ultramicro Leu-leu endopeptid ase in buccal ganglion of Aplysia California by MALDI-TOF mass spectrometry.

利用丙酮 -水 -酸 (4 8∶ 5 0∶ 2 V/ V)萃取系统 ,从加州海兔 (Aplysia California,AC)口腔神经节中提取 L eu- L eu多肽酶 ,并用反相 HPL C进行纯化。选用具有三对 L eu- L eu结构的酸性多肽 (acidic peptide,AP)作为探针 ,以基质辅助激光解吸电离飞行时间 (MAL DI- TOF)质谱技术研究了海兔口腔神经节中的超微量 L eu- L eu多肽酶。对 L eu- L eu多肽酶的活性与类型及温度处理效果进行讨论。

Leu-Leu endopeptidase in buccal ganglion (BG) of Aplysia California (AC) was extracted with acetone- water-HCl (48∶50∶2) system and purified by RP-HPLC. Using acidic peptide with three pairs of Leu-Leu structure as probe , the ultramicro Leu-Leu endopeptidase in BG of AC was analyzed by MALDI-TOF-MS. The activities and kinds of Leu-Leu endopeptidase and the effect of temperature treatment are discussed. The technique is also applicable for analyzing ultramicro endopeptidases in other tissues or cells.