Chougule Nanasaheb P, Giri Ashok P, Sainani Mohini N, Gupta Vidya S
National Chemical Laboratory, Plant Molecular Biology Unit, Division of Biochemical Sciences, Dr. Homi Bhabha Road, Pune 411 008 (MS), India.
Insect Biochem Mol Biol. 2005 Apr;35(4):355-67. doi: 10.1016/j.ibmb.2005.01.006.
Relative quantification of reported gut proteinase cDNAs from Helicoverpa armigera larvae fed on various host plants (cotton, chickpea, pigeonpea, tomato and okra), non-host plant PIs (winged bean, bitter gourd, ground nut, and capsicum) and during larval development has been carried out using semi-quantitative RT-PCR. Five trypsin-like and three chymotrypsin-like proteinases were categorized as insensitive or sensitive to most of the proteinase inhibitors (PIs) and insensitive/sensitive to specific PIs based on their expression analysis. These results were supported by amino acid sequence analysis, where diverged amino acids were observed in the regions, which are reported to be involved in typical trypsin-trypsin inhibitor interactions and critical for proteinase inhibitor resistance. Among exopeptidases (five aminopeptidase and three carboxypeptidase), HaAmi4 and HaAmi5 of aminopeptidase and HaCar1 of carboxypeptidase exhibited considerable differential expression. Elastase and cathepsin B-like proteinases were expressed at negligible levels. The proteases identified in the study would be ideal candidates for further interactions studies with protease inhibitors to understand the structural reasons of protease inhibitor insensitivity.
利用半定量逆转录聚合酶链反应(RT-PCR),对取食不同寄主植物(棉花、鹰嘴豆、木豆、番茄和秋葵)、非寄主植物蛋白酶抑制剂(四棱豆、苦瓜、花生和辣椒)的棉铃虫幼虫以及幼虫发育过程中已报道的肠道蛋白酶cDNA进行了相对定量分析。根据表达分析,5种类胰蛋白酶和3种类胰凝乳蛋白酶被归类为对大多数蛋白酶抑制剂(PIs)不敏感或敏感,以及对特定PIs不敏感/敏感。这些结果得到了氨基酸序列分析的支持,在据报道参与典型胰蛋白酶-胰蛋白酶抑制剂相互作用且对蛋白酶抑制剂抗性至关重要的区域观察到了不同的氨基酸。在外肽酶(5种氨肽酶和3种羧肽酶)中,氨肽酶的HaAmi4和HaAmi以及羧肽酶的HaCar1表现出相当大的差异表达。弹性蛋白酶和组织蛋白酶B样蛋白酶的表达水平可忽略不计。该研究中鉴定出的蛋白酶将是进一步与蛋白酶抑制剂进行相互作用研究的理想候选对象,以了解蛋白酶抑制剂不敏感的结构原因。