Response of Midgut Trypsin- and Chymotrypsin-Like Proteases of Helicoverpa armigera Larvae Upon Feeding With Peanut BBI: Biochemical and Biophysical Characterization of PnBBI
| Autor: | Marri Swathi, Vadthya Lokya, Nalini Mallikarjuna, Kollipara Padmasree |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
0301 basic medicine Proteases medicine.medical_treatment two-dimensional zymography Plant Science PnBBI Helicoverpa armigera (Noctuidae) Helicoverpa armigera lcsh:Plant culture 01 natural sciences 03 medical and health sciences medicine Zymography lcsh:SB1-1110 Protease Chymotrypsin biology Molecular mass Chemistry fungi Midgut Trypsin biology.organism_classification circular dichroism 030104 developmental biology Biochemistry biology.protein Arachis hypogaea (Fabaceae) surface plasmon resonance 010606 plant biology & botany medicine.drug |
| Zdroj: | Frontiers in Plant Science, Vol 11 (2020) |
| DOI: | 10.3389/fpls.2020.00266/full |
| Popis: | Proteinase/Protease inhibitors (PIs) from higher plants play an important role in defense and confer resistance against various insect pests and pathogens. In the present study, Bowman-Birk Inhibitor (BBI) was purified from mature seeds of an interspecific advanced hybrid peanut variety (4368-1) using chromatographic techniques. The biochemical and biophysical characteristics such as low molecular mass, presence of several isoinhibitors and higher-ordered dimer/tetramer, predominance of antiparallel β-sheets and random coils in secondary structure, reactive sites against trypsin and chymotrypsin, broad spectrum of stability toward extreme pH and temperature along with MALDI TOF-TOF analysis (ProteomeXchange identifier PXD016933) ascertained the purified biomolecule from peanut as BBI (PnBBI). Surface plasmon resonance competitive binding analysis revealed the bifunctional PnBBI is a trypsin specific inhibitor with 1:2 stoichiometry as compared to chymotrypsin. A concentration-dependent self-association tendency of PnBBI was further confirmed by 'red shift' in the far-UV CD spectra. Furthermore, the insecticidal potential of PnBBI against Helicoverpa armigera was assessed by in vitro assays and in vivo feeding experiments. A significant reduction in larval body weight was observed with concomitant attenuation in the activity of midgut trypsin-like proteases of H. armigera (HaTPs) fed on PnBBI supplemented diet. The one and two-dimensional zymography studies revealed the disappearance of several isoforms of HaTP upon feeding with PnBBI. qRT-PCR analysis further suggests the role of PnBBI in not only inhibiting the activity of midgut trypsin and chymotrypsin-like proteases but also in modulating their expression. Taken together, the results provide a biochemical and molecular basis for introgressed resistance in peanut interspecific advanced hybrid variety against H. armigera. |
| Databáze: | OpenAIRE |
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