Interaction of almond cystatin with pesticides: Structural and functional analysis

Abstract

Pesticides are chemical substances that eliminate or control a variety of agricultural pests that damage crops and livestock. They not only affect the targeted pests but also affect the nontargeted systems, raising more concerns for their effect on both plant and animal systems. Cystatins (cysteine protease inhibitor) are ubiquitously present in all living cells and show a variety of important physiological functions. The present study shows the effect of different pesticides (pendimethalin, methoxyfenozide, and Cu hydroxide) on purified almond cystatin. Almond cystatin showed concentration-dependent loss in papain inhibitory activity on interaction with the pesticides, showing maximum loss in the presence of Cu(II) hydroxide and minimum in the case of methoxyfenozide. Native polyacrylamide gel electrophoresis showed maximum degradation of purified cystatin in the presence of Cu(II) hydroxide with insignificant effect in the presence of methoxyfenozide. Structural alterations were significant in the case of Cu(II) hydroxide and less in the case of methoxyfenozide as revealed by UV and fluorescence spectral studies. Secondary structural alterations were further conformed by circular dichroism and Fourier transform infrared spectroscopy. The α-helix content of almond cystatin decreases from 35.64% (native) to 34.83%, 30.79%, and 29.62% for methoxyfenozide-, pendimethalin-, and Cu(II) hydroxide-treated cystatin, respectively. A Fourier transform infrared study shows an amide I band shift for almond cystatin from 1649.15 ± 0.5 to 1646.48 ± 0.6, 1640.44 ± 0.6, and 1635.11 ± 0.3 cm for methoxyfenozide, pendimethalin, and Cu(II) hydroxide, respectively. Values obtained for different thermodynamic parameters (ΔH , ΔG , N, and ΔS ) by isothermal titration calorimetric experiments reveal maximum binding of almond cystatin with Cu(II) hydroxide followed by pendimethalin and little interaction with methoxyfenozide.

Genome-wide identification and analysis of cystatin family genes in Sorghum (Sorghum bicolor (L.) Moench)

“…Plant cystatins are involved in various biotic stress responses and probably act as defense proteins against pest infestation and pathogen infection ( Meriem et al, 2010 ). At present, some cystatins with insecticidal activity have been isolated from many plants, such as barley, tomato, and potato ( Rasoolizadeh et al, 2017 ; Siddiqui, Khaki & Bano, 2017 ; Velasco-Arroyo et al, 2018 ; Goulet, Sainsbury & Michaud, 2020 ). Several cystatins having antifungal activities were also isolated from taro, cacao, and wheat ( Christova et al, 2018 ; Pirovani et al, 2010 ; Chen et al, 2014 ).…”

Peer Review #4 of "Genome-wide identification and analysis of cystatin family genes in Sorghum (Sorghum bicolor (L.) Moench) (v0.2)"

“…Manuscript to be reviewed Plant cystatins are involved in various biotic stress responses and probably act as defense proteins against pest infestation and pathogen infection (Meriem et al 2010). At present, some cystatins with insecticidal activity have been isolated from many plants, such as barley, tomato, and potato (Rasoolizadeh et al 2017;Siddiqui et al 2017;Velasco-Arroyo et al 2018;Goulet et al 2020). Several cystatins having antifungal activities were also isolated from taro, cacao, and wheat (Christova et al 2018;Pirovani et al 2010;Chen et al 2014).…”