›› 2001, Vol. 44 ›› Issue (1): 51-55.

• 研究论文 • 上一篇    下一篇

家蚕滞育性卵盐酸处理的靶物质

徐世清1,郑必平1,司马杨虎1,甲斐英则2,徐俊良3   

  • 出版日期:2001-02-20 发布日期:2001-02-20

Target of hydrochloric acid in acid-treatment of diapausing eggs of Bombyx mori

XU Shi-qing,ZHENG Bi-ping,SIMA Yang-hu,Kai Hideno-ri2,XU Jun-liang3   

  • Online:2001-02-20 Published:2001-02-20

摘要: 酯酶A4(EA4)是家蚕卵的滞育生物钟蛋白质。从家蚕C108品种产下后48 h的滞育性卵和盐酸活化处理卵分离纯化出EA4酶蛋白,使用合成的EA4活性多肽抑制因子PIN(氨基酸结构:SIFMTKQHSQ DDIIQHPLDY VEQQIHQQKQ KLQKQTLN),研究了PIN对EA4酶蛋白的作用机制。滞育性卵的EA4酶蛋白和PIN在25℃混合24h后,用矩阵辅助激光解吸离子质谱法,检测到了二者的结合体,该结合体在盐酸处理后消失;盐酸活化处理蚕卵的EA4酶蛋白和合成PIN之间没有出现这种结合体。体外25℃,滞育性蚕卵EA4的ATPase特征性活性峰在6.5 h后出现,而盐酸活化处理蚕卵的EA4在1.5 h后出现活性峰值。盐酸处理可能通过解除PIN对EA4的抑制作用,在短时间内激活EA4酶蛋白,从而活化滞育性蚕卵。

关键词: 家蚕, 滞育, 盐酸处理, 酯酶A4, 靶物质

Abstract: Esterase A4(EA4), a time interval measuring enzyme in the diapausing eggs of Bombyx mori, was isolated and purified from C108 silkworm diapausing eggs and acid-treated eggs 2 days after oviposition, respectively. The PIN is a kind of peptide inhibitory needle for EA4 enzyme activity. Its amino acid sequence is SIFMTKQHSQ DDIIQHPLDY VEQQIHQQKQ KLQKQTLN. The action of PIN on EA4 was studied in the paper. When the enzymic protein EA4 from diapausing eggs was incubated with the synthetic PIN for 24 h at 25℃, a complex of EA4 and PIN was detected by matrix-assisted laser desorption ionization mass spectrometry. When the treated mixture above was incubated again in 2.5% HCl, the complex could not be found. There is no complex formed when the EA4 protein from acid-treated eggs was incubated with the synthetic PIN. At 25℃ in vitro, the activity peak of EA4 from diapausing eggs and acid-treated eggs for ATPase appeared at 6.5 h and 1.5 h, respectively. The results showed that the activation of diapausing eggs was due to eliminating the inhibition of PIN on EA4 and activating EA4 by HCl within a short time.

Key words: Bombyx mori, diapause, hydrochloric acid\|treatment, esterase A4, target