Vegetos
(An International Journal of Plant Research & Biotechnology)
Structural elucidation of Cocculus hirsutus trypsin inhibitor and insights into its mechanism of action
*Article not assigned to an issue yet
Research Articles | Published: 07 May, 2025
First Page: 0
Last Page: 0
Views: 332
Keywords: In silico docking, n Cocculus hirsutus trypsin inhibitor, Trypsin, Protein interaction, Molecular dynamic stimulation
Abstract
Many plants are known to contain proteinase inhibitors in their vegetative parts and seeds, which play important role as defence molecules to overcome biotic and abiotic stresses. Cocculus hirsutus trypsin inhibitor (ChTI), isolated from leaves of Cocculus hirsutus is characterized to be stable protein at temperature 70 °C and pH 7–9. In vitro experiments show that ChTI inhibits trypsin and exhibits deleterious effect on growth and development of lepidopteron insects that have trypsin like gut proteinase activity. However, these insects successfully adapt to proteinase inhibitors of their host plants by switching the expression of proteinase genes. Thus, it is interesting to study the interaction between proteinase inhibitors from non-host plant and gut proteinases. In silico interaction of trypsin was carried out using the predicted three dimensional structure of ChTI. Docking studies suggest that ChTI is a non canonical type of inhibitor and form stable inhibitor complexes with Trypsin and Chymotrypsin.
References
Bateman KS, James MN (2011) Plant protein proteinase inhibitors: Structure and mechanism of inhibition. Curr Protein Pept Sci 12:340–347
Bendre AD, Suresh CG, Shanmugam D, Sureshkumar R (2018) Structural insights into the unique inhibitory mechanism of Kunitz type trypsin inhibitor from Cicer arietinum L. J Biomol Struct Dyn 37:2669–2677
Bhattacherjee C, Manjunath NH, Prasad DT (2010) Purification of a trypsin inhibitor from Cocculus hirsutus and identification of its biological activity. J Crop Sci Biotech 12:253–260
Buchan DWA, Jones DT (2019) The PSIPRED protein analysis workbench: 20 years on. Nucleic Acids Res 47:402–407
Chattopadhyay P, Banerjee G, Mukherjee S (2017) Recent trends of modern bacterial insecticides for pest control practice in integrated crop management system. 3 Biotech 7:60
Cui X, Du J, Li J, Wang Z (2018) Inhibitory site of α-hairpinin peptide from tartary buckwheat has no effect on its antimicrobial activities. Acta Biochim Biophys Sin 50:408–416
DeLano WL (2002) Pymol: An open-source molecular graphics tool. CCP4 Newslett Protein Crystallogr 40:82–92
Dhaliwal GS, Jindal V, Mohindru B (2015) Crop losses due to insect pests: Global and Indian scenario. Indian J Ecol 77:165–168
Dias RO, Via A, Brandao MM, Tramontano A, Silva-Filho MC (2015) Digestive peptidase evolution in holometabolous insects led to a divergent group of enzymes in Lepidoptera. Insect Biochem Mol Biol 58:1–11
Drozdetskiy A, Cole C, Procter J, Barton GJ (2015) J Pred 4: a protein secondary structure prediction server. Nucleic Acids Res 43:389–394
Dunse K, Stevens J, Lay F, Gaspar Y, Heath R, Anderson M (2010) Co-expression of potato type I and II proteinase inhibitors gives cotton plants protection against insect damage in the field. Proc Natl Acad Sci USA 107:15011–15015
Kolb P, Ferreira RS, Irwin JJ, Shoichet BK (2009) Docking and chemo-informatic screens for new ligands and targets. Cur Opin Biotechnol 20:429–436
Kozakov D, Hall DR, Xia B, Porter KA, Padhorny D, Yueh C, Beglov D, Vajda S (2017) The ClusPro web server for protein-protein docking. Nat Protoc 12:255–278
Kriticos DJ, Ota N, Hutchison WD, Beddow J, Walsh T (2015) The potential distribution of invading Heicoverpa armigera in North America: is just a matter of time? PLoS ONE 10:1–10
Laskowski RA, Swindells MB (2011) LigPlot+: multiple ligand-protein interaction diagrams for drug discovery. J Chem Inf Model 2011(51):2778–2786
Laskowski RA, MacArthur MW, Moss DS, Thornton JM (1993) PROCHECK: a program to check the stereochemical quality of protein structures. J Appl Crystallogr 26:283–291
Le Goff G, Giraudo M (2019) Effects of pesticides on the environment and insecticide resistance. Olfactory concepts of insect control-alternative to insecticides. Springer, Heidelberg, pp 51–78
Liu Z, Zhu Q, Li J, Zhang G, Jiamahate A, Zhou J, Liao H (2015) Isolation, structure modeling and function characterization of a trypsin inhibitor from Cassia obtusifolia. Biotechnol Lett 37:863–869
Manushree V, Devaraj VR, Prasad DT (2020) Expression of Cocculus hirsutus trypsin inhibitor promotes endogenous defensive response against Helicoverpa armigera and enhanced levels of antioxidants. African J Plant Sci 14:265–282
Notredame C, Higgins DG, Heringa J (2000) T-Coffee: a novel method for fast and accurate multiple sequence alignment. J Mol Biol 302:205–217
Oliva ML, Sampaio MU (2009) Action of plant proteinase inhibitors on enzymes of physio pathological importance. An Acad Bras Cienc 81:615–621
Oliveira MG, Rogana E, Rosa JC, Reinhold BB, Andrade MH, Greene LJ, Mares-Guia M (1993) Tyrosine 151 is part of the substrate activation binding site of bovine trypsin. Identification by covalent labeling with p-diazoniumbenzamidine and kinetic characterization of Tyr-151-(p-benzamidino)-azo-beta-trypsin. J Biol Chem 268:26893–26903
Oparin PB, Mineev KS, Dunaevsky YE, Arseneiv AS, Belozersky MA, Grishin EV, Egorov TA, Vassilevski AA (2012) Buckwheat trypsin inhibitor with helical hairpin structure belongs to a new family of plant defence peptides. Biochem J 446:69–77
Pang Z, Zhou Z, Yin D (2013) Transgenic rice plants overexpressing BBTI4 confer partial but broad-spectrum bacterial blight resistance. J Plant Biol 56:383
Renko M, Sabotic J, Turk D (2012) β-Trefoil inhibitors from the work of Kunitz onward. Biol Chem 393:1043–1054
Sievers F, Higgins DG (2018) Clustal Omega for making accurate alignments of many protein sciences. Protein Sci 27:135–145
Souza TP, Dias RO, Castelhano EC, Brandao MM, Moura DS, Silva-Filho MC (2016) Comparative analysis of expression profiling of the trypsin and chymotrypsin genes from Lepidoptera species with different levels of sensitivity to soybean peptidase inhibitors. Comp Biochem Physiol B Biochem Mol Biol 196:67–73
Srinivasan A, Giri AP, Vidya SG (2006) Structural and functional diversities in lepidopteran serine proteases. Cell Mol Biol Lett 11:132–154
Swathi M, Mishra PK, Lokya V, Swaroop V, Mallikarjuna N, Dutta-Gupta A, Padmasree K (2016) Purification and partial characterization of trypsin-specific proteinase inhibitors from pigeon pea wild relative Cajanus platycarpus L. (Fabaceae) active against gut proteases of lepidopteran pest Helicoverpa armigera. Front Physiol 7:388
Tanpure RS, Barbole RS, Dawkar VV, Waichal AY, Joshi RS, Giri AP, Gupta VS (2017) Improved tolerance against Helicoverpa armigera in transgenic tomato over-expressing multi-domain proteinase inhibitor gene from Capsicum annuum. Physiol Mol Biol Plants 23:597–604
Vijravijayan S, Plentnev S, Nandhagopal VZ, Gunasekaran K (2018) Crystal structure of a novel Kunitz type inhibitor, alocasin with anti-aedesaegypti activity targeting midgut proteases. Pest Manag Sci 74:2761–2772
Xu D, Zhang D (2011) Improving the physical realism and structural accuracy of protein models by a two-step atomic-level energy minimization. Biophys J 101:2525–2534
Yang L, Manithody C, Rezaie AR (2007) The role of autolysis loop in determining the specificity of coagulation proteases. Braz J Med Biol Res 40:1055–1064
Yang J, Yan R, Roy A, Xu D, Poisson J, Zhang Y (2015) The I-TASSER suite: protein structure and function prediction. Nat Methods 12:7–8
Zhou D, Lobo YA, Batista IFC, Marques- Porto R, Gustchina A, OlivaWlodawer MLVA (2013) Crystal structures of a plant trypsin inhibitor from Enterolobium contortisiliquum (EcTI) and of its complex with bovine trypsin. PLoS ONE 8:1–15
Tyndall TDA, Nall T, Fairlie DP (2005) Proteases universally recognize beta strands in their active sites. Chem Rev 105:973–999
Rustgi S, Fontvieille EB, Reinbothe C, Wettstein DV, Reinbothe S (2018) The complex world of plant protease inhibitors: Insights into a Kunitz-type cysteine protease inhibitor of Arabidopsis thaliana. Commun Integr Biol 11(1):e1368599
De Oliveira CF, Oliveira TMF, Cardoso MH, Nogueira KG, Russi R, De França AF, Dos Santos EA, Franco OL, De Oliveira AS Migliolo L (2019) Dual insecticidal effects of Adenanthera pavonina kunitz-type inhibitor on Plodiainter punctella is mediated by digestive enzymes inhibition and chitin-binding properties. Molecules 24:4344
Chen X, Riley BT, Veer SJD, Hoke DE, Haeften DE, Leahy D, Harris JM (2019) Potent multi-target serine protease inhibition achieved by a simplified β-sheet motif. PloS one 14:1–15
Author Information
Department of Biochemistry, Bangalore City University, Bangalore, India