Inhibitory specificity and insecticidal selectivity of ??-amylase inhibitor from Phaseolus vulgaris

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• 作者：Kluh ; Ivan ; Horn ; Martin ; H??blov?? ; Jana ; Hubert ; Jan ; Dole??kov??-Mare??ov?? ; Lucie ; et. al.
• 关键词：????AI-1 ; ??-Amylase inhibitor from common bean (Phaseolus vulgaris) ; BPTI ; bovine pancreatic trypsin inhibitor ; PDB ; Protein Data Bank ; RP-HPLC ; reverse-phase high performance liquid chromatography ; SDS??????PAGE ; sodium dodecyl sulfat
• 刊名：Phytochemistry
• 出版年：2005
• 出版时间：January, 2005
• 年：2005
• 卷：66
• 期：1
• 页码：31-39
• 全文大小：473 K

文摘

The primary structure and proteolytic processing of the ??-amylase isoinhibitor ?? AI-1 from common bean (Phaseolus vulgaris cv. Magna) was determined by protein chemistry techniques. The inhibitory specificity of ??AI-1 was screened with a panel of the digestive ??-amylases from 30 species of insects, mites, gastropod, annelid worm, nematode and fungal phytopathogens with a focus on agricultural pests and important model species. This in vitro analysis showed a selective inhibition of ??-amylases from three orders of insect (Coleoptera, Hymenoptera and Diptera) and an inhibition of ??-amylases of the annelid worm. The inhibitory potential of ??AI-1 against several ??-amylases was found to be modulated by pH. To understand how ??AI-1 discriminates among closely related ??-amylases, the sequences of the ??-amylases sensitive, respectively, insensitive to ??AI-1 were compared, and the critical determinants were localized on the spatial ??-amylase model. Based on the in vitro analysis of the inhibitory specificity of ??AI-1, the in vivo activity of the ingested ??AI-1 was demonstrated by suppression of the development of the insect larvae that expressed the sensitive digestive ??-amylases. The first comprehensive mapping of ??AI-1 specificity significantly broadens the spectrum of targets that can be regulated by ??-amylase inhibitors of plant origin, and points to potential application of these protein insecticides in plant biotechnologies.