家蚕滞育性卵盐酸处理的靶物质  

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

作　　者：徐世清[1] 郑必平[1] 司马杨虎[1] 甲斐英则[2] 徐俊良[3] 

机构地区：[1]苏州大学蚕丝生物技术实验室,苏州215151 [2]日本国立鸟取大学农学部 [3]浙江大学蚕学系,杭州310029

出　　处：《昆虫学报》2001年第1期51-55,共5页Acta Entomologica Sinica

摘　　要：酯酶A4 (EA4 )是家蚕卵的滞育生物钟蛋白质。从家蚕C10 8品种产下后 4 8h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 :SIFMTKQHSQDDIIQHPLDYVEQQIHQQKQKLQKQTLN) ,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 2 5℃混合 2 4h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 2 5℃ ,滞育性蚕卵EA4的ATPase特征性活性峰在 6 5h后出现 ,而盐酸活化处理蚕卵的EA4在 1 5h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。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.

关 键 词：家蚕 滞育 盐酸处理 酯酶A4 靶物质 卵 

分 类 号：S882.4[农业科学—特种经济动物饲养]