Silva C P., Terra W R., de Sá M F.G., Samuels R I., Isejima E M., Bifano T D., Almeida J S.
Laboratório de Qui;mica e Função de Protei;nas e Pepti;deos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, CEP 28015-620, Campos dos Goytacazes, Brazil
J Insect Physiol. 2001 Nov;47(11):1283-1290. doi: 10.1016/s0022-1910(01)00115-9.
Zabrotes subfasciatus larvae possess three alpha-amylase isoforms determined by in gel assays following SDS-PAGE. Two minor isoforms present lower electrophoretic mobility than the major form. When developed inside Vigna unguiculata (cowpea) seeds, fourth instar larvae have minor quantities of the slow-migrating isoforms, but when reared on seeds of Phaseolus vulgaris (common bean), the two slow-migrating forms are expressed in higher amounts, whilst the quantity of the major constitutive form is independent of the host bean. Larvae at the beginning of the fourth instar were fed on flour or cotyledons of cowpea and common bean and it was observed that the larvae fed on the common bean expressed the two slow-migrating forms in higher amounts when compared to the control larvae fed on cowpea. In order to investigate the possible correlation between the induction of alpha-amylases and the ingestion of the common bean alpha-amylase inhibitor 1 (alphaAI-1), this inhibitor was incorporated into artificial diet. It was observed that larvae fed on diet containing chronic doses of alphaAI-1 during their development, produced the two slow-migrating forms in higher amounts than control larvae, however, fourth-instar larvae fed on the same diet presented less amylase activity than control larvae. The data suggested that alphaAI-1 is involved in amylase induction and that it has inhibitory activity against the constitutive amylase, when starch granules are used as substrate.
豆象幼虫拥有三种α-淀粉酶同工型,这是通过SDS-PAGE后的凝胶分析确定的。两种次要同工型的电泳迁移率低于主要同工型。在豇豆种子内发育时,四龄幼虫体内的慢速迁移同工型含量较少,但在菜豆种子上饲养时,两种慢速迁移形式的表达量更高,而主要组成型同工型的含量与宿主豆无关。给四龄初期的幼虫喂食豇豆和菜豆的面粉或子叶,观察到与以豇豆为食的对照幼虫相比,以菜豆为食的幼虫中两种慢速迁移形式的表达量更高。为了研究α-淀粉酶的诱导与菜豆α-淀粉酶抑制剂1(αAI-1)摄入之间的可能相关性,将这种抑制剂添加到人工饲料中。观察到在发育过程中以含有慢性剂量αAI-1的饲料为食的幼虫,产生的两种慢速迁移形式的量比对照幼虫多,然而,以相同饲料为食的四龄幼虫的淀粉酶活性比对照幼虫低。数据表明,αAI-1参与淀粉酶诱导,并且当淀粉颗粒用作底物时,它对组成型淀粉酶具有抑制活性。