Crystal structure of Kunitz-type trypsin inhibitor: Entomotoxic effect of native and encapsulated protein targeting gut trypsin of Tribolium castaneum Herbst

Sohaib Mehmood; Soren Skou Thirup; Sarah Ahmed; Nabila Bashir; Ahsan Saeed; Maria Rafiq; Qamar Saeed; Muhammad Najam-ul-Haq; Binish Khaliq; Muhammad Ibrahim; Wadi Brak Alonazi; Ahmed Akrem

doi:10.1016/j.csbj.2024.07.023

Crystal structure of Kunitz-type trypsin inhibitor: Entomotoxic effect of native and encapsulated protein targeting gut trypsin of Tribolium castaneum Herbst

Sohaib Mehmood, Soren Skou Thirup, Sarah Ahmed, Nabila Bashir, Ahsan Saeed, Maria Rafiq, Qamar Saeed, Muhammad Najam-ul-Haq, Binish Khaliq, Muhammad Ibrahim, Wadi Brak Alonazi, Ahmed Akrem^*

^*Corresponding author for this work

Department of Molecular Biology and Genetics - Protein Science

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

Abstract

Trypsin inhibitors are known to act against insect pests by inhibiting proteases of the digestive tract. In this study, we report structural and functional characterization of ∼ 19 kDa Albizia procera Kunitz-type trypsin inhibitor (ApKTI) protein with potential bio-insecticidal applications. Crystal structure of ApKTI protein has been refined to 1.42 Å and molecular structure (8HNR) showed highly beta sheeted conformation including 12 beta sheets, 15 loops and two small alpha helices. Docking between predicted model of Tribolium castaneum trypsin (TcPT) and 8HNR produced a stable complex (−11.3 kcal/mol) which reflects the inhibitory potential of ApKTI against insect gut trypsin. Significant mortality was observed in all life stages of T. castaneum including egg, larvae, pupae and adults with a 3.0 mg native ApKTI treatment in comparison to negative control. Although standard trypsin inhibitor (Glycine max trypsin inhibitors; GmKTI; 3.0 mg) produced maximum reduction against all above life stages; however, a non-significant mortality difference was observed in comparison to 3.0 mg native ApKTI. The study further explores the synthesis and characterization of Graphene (GNPs) and Zinc oxide (ZnONPs) nanoparticles, followed by the optimization of ApKTI and GmKTI loading on both nanoparticles to evaluate their enhanced insecticidal effectiveness. Encapsulated proteins showed significant mortality against T. castaneum across all concentrations, with GNPs proving more effective than ZnONPs. Additionally, encapsulated GmKTI produced significant mortality of eggs compared to loaded ApKTI treatments while other life stages were non-significantly affected by two proteins. This research highlights the importance of encapsulated ApKTI protein for eco-friendly pest management strategies.

Original language	English
Journal	Computational and Structural Biotechnology Journal
Volume	23
Pages (from-to)	3132-3142
Number of pages	11
ISSN	2001-0370
DOIs	https://doi.org/10.1016/j.csbj.2024.07.023
Publication status	Published - Dec 2024

Keywords

Albizia procera
Crystal structure
Graphene nanoparticles
Kunitz–type trypsin inhibitor
Zinc oxide nanoparticles

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10.1016/j.csbj.2024.07.023Licence: CC BY-NC-ND

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Mehmood, S., Thirup, S. S., Ahmed, S., Bashir, N., Saeed, A., Rafiq, M., Saeed, Q., Najam-ul-Haq, M., Khaliq, B., Ibrahim, M., Alonazi, W. B., & Akrem, A. (2024). Crystal structure of Kunitz-type trypsin inhibitor: Entomotoxic effect of native and encapsulated protein targeting gut trypsin of Tribolium castaneum Herbst. Computational and Structural Biotechnology Journal, 23, 3132-3142. https://doi.org/10.1016/j.csbj.2024.07.023

@article{b2ade19547b5468ca3841bdf3990d475,

title = "Crystal structure of Kunitz-type trypsin inhibitor: Entomotoxic effect of native and encapsulated protein targeting gut trypsin of Tribolium castaneum Herbst",

abstract = "Trypsin inhibitors are known to act against insect pests by inhibiting proteases of the digestive tract. In this study, we report structural and functional characterization of ∼ 19 kDa Albizia procera Kunitz-type trypsin inhibitor (ApKTI) protein with potential bio-insecticidal applications. Crystal structure of ApKTI protein has been refined to 1.42 {\AA} and molecular structure (8HNR) showed highly beta sheeted conformation including 12 beta sheets, 15 loops and two small alpha helices. Docking between predicted model of Tribolium castaneum trypsin (TcPT) and 8HNR produced a stable complex (−11.3 kcal/mol) which reflects the inhibitory potential of ApKTI against insect gut trypsin. Significant mortality was observed in all life stages of T. castaneum including egg, larvae, pupae and adults with a 3.0 mg native ApKTI treatment in comparison to negative control. Although standard trypsin inhibitor (Glycine max trypsin inhibitors; GmKTI; 3.0 mg) produced maximum reduction against all above life stages; however, a non-significant mortality difference was observed in comparison to 3.0 mg native ApKTI. The study further explores the synthesis and characterization of Graphene (GNPs) and Zinc oxide (ZnONPs) nanoparticles, followed by the optimization of ApKTI and GmKTI loading on both nanoparticles to evaluate their enhanced insecticidal effectiveness. Encapsulated proteins showed significant mortality against T. castaneum across all concentrations, with GNPs proving more effective than ZnONPs. Additionally, encapsulated GmKTI produced significant mortality of eggs compared to loaded ApKTI treatments while other life stages were non-significantly affected by two proteins. This research highlights the importance of encapsulated ApKTI protein for eco-friendly pest management strategies.",

keywords = "Albizia procera, Crystal structure, Graphene nanoparticles, Kunitz–type trypsin inhibitor, Zinc oxide nanoparticles",

author = "Sohaib Mehmood and Thirup, {Soren Skou} and Sarah Ahmed and Nabila Bashir and Ahsan Saeed and Maria Rafiq and Qamar Saeed and Muhammad Najam-ul-Haq and Binish Khaliq and Muhammad Ibrahim and Alonazi, {Wadi Brak} and Ahmed Akrem",

note = "Publisher Copyright: {\textcopyright} 2024 The Authors",

year = "2024",

month = dec,

doi = "10.1016/j.csbj.2024.07.023",

language = "English",

volume = "23",

pages = "3132--3142",

journal = "Computational and Structural Biotechnology Journal",

issn = "2001-0370",

publisher = "Research Network of Computational and Structural Biotechnology (RNCSB)",

}

Mehmood, S, Thirup, SS, Ahmed, S, Bashir, N, Saeed, A, Rafiq, M, Saeed, Q, Najam-ul-Haq, M, Khaliq, B, Ibrahim, M, Alonazi, WB & Akrem, A 2024, 'Crystal structure of Kunitz-type trypsin inhibitor: Entomotoxic effect of native and encapsulated protein targeting gut trypsin of Tribolium castaneum Herbst', Computational and Structural Biotechnology Journal, vol. 23, pp. 3132-3142. https://doi.org/10.1016/j.csbj.2024.07.023

Crystal structure of Kunitz-type trypsin inhibitor: Entomotoxic effect of native and encapsulated protein targeting gut trypsin of Tribolium castaneum Herbst. / Mehmood, Sohaib; Thirup, Soren Skou; Ahmed, Sarah et al.
In: Computational and Structural Biotechnology Journal, Vol. 23, 12.2024, p. 3132-3142.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

TY - JOUR

T1 - Crystal structure of Kunitz-type trypsin inhibitor

T2 - Entomotoxic effect of native and encapsulated protein targeting gut trypsin of Tribolium castaneum Herbst

AU - Mehmood, Sohaib

AU - Thirup, Soren Skou

AU - Ahmed, Sarah

AU - Bashir, Nabila

AU - Saeed, Ahsan

AU - Rafiq, Maria

AU - Saeed, Qamar

AU - Najam-ul-Haq, Muhammad

AU - Khaliq, Binish

AU - Ibrahim, Muhammad

AU - Alonazi, Wadi Brak

AU - Akrem, Ahmed

PY - 2024/12

Y1 - 2024/12

N2 - Trypsin inhibitors are known to act against insect pests by inhibiting proteases of the digestive tract. In this study, we report structural and functional characterization of ∼ 19 kDa Albizia procera Kunitz-type trypsin inhibitor (ApKTI) protein with potential bio-insecticidal applications. Crystal structure of ApKTI protein has been refined to 1.42 Å and molecular structure (8HNR) showed highly beta sheeted conformation including 12 beta sheets, 15 loops and two small alpha helices. Docking between predicted model of Tribolium castaneum trypsin (TcPT) and 8HNR produced a stable complex (−11.3 kcal/mol) which reflects the inhibitory potential of ApKTI against insect gut trypsin. Significant mortality was observed in all life stages of T. castaneum including egg, larvae, pupae and adults with a 3.0 mg native ApKTI treatment in comparison to negative control. Although standard trypsin inhibitor (Glycine max trypsin inhibitors; GmKTI; 3.0 mg) produced maximum reduction against all above life stages; however, a non-significant mortality difference was observed in comparison to 3.0 mg native ApKTI. The study further explores the synthesis and characterization of Graphene (GNPs) and Zinc oxide (ZnONPs) nanoparticles, followed by the optimization of ApKTI and GmKTI loading on both nanoparticles to evaluate their enhanced insecticidal effectiveness. Encapsulated proteins showed significant mortality against T. castaneum across all concentrations, with GNPs proving more effective than ZnONPs. Additionally, encapsulated GmKTI produced significant mortality of eggs compared to loaded ApKTI treatments while other life stages were non-significantly affected by two proteins. This research highlights the importance of encapsulated ApKTI protein for eco-friendly pest management strategies.

AB - Trypsin inhibitors are known to act against insect pests by inhibiting proteases of the digestive tract. In this study, we report structural and functional characterization of ∼ 19 kDa Albizia procera Kunitz-type trypsin inhibitor (ApKTI) protein with potential bio-insecticidal applications. Crystal structure of ApKTI protein has been refined to 1.42 Å and molecular structure (8HNR) showed highly beta sheeted conformation including 12 beta sheets, 15 loops and two small alpha helices. Docking between predicted model of Tribolium castaneum trypsin (TcPT) and 8HNR produced a stable complex (−11.3 kcal/mol) which reflects the inhibitory potential of ApKTI against insect gut trypsin. Significant mortality was observed in all life stages of T. castaneum including egg, larvae, pupae and adults with a 3.0 mg native ApKTI treatment in comparison to negative control. Although standard trypsin inhibitor (Glycine max trypsin inhibitors; GmKTI; 3.0 mg) produced maximum reduction against all above life stages; however, a non-significant mortality difference was observed in comparison to 3.0 mg native ApKTI. The study further explores the synthesis and characterization of Graphene (GNPs) and Zinc oxide (ZnONPs) nanoparticles, followed by the optimization of ApKTI and GmKTI loading on both nanoparticles to evaluate their enhanced insecticidal effectiveness. Encapsulated proteins showed significant mortality against T. castaneum across all concentrations, with GNPs proving more effective than ZnONPs. Additionally, encapsulated GmKTI produced significant mortality of eggs compared to loaded ApKTI treatments while other life stages were non-significantly affected by two proteins. This research highlights the importance of encapsulated ApKTI protein for eco-friendly pest management strategies.

KW - Albizia procera

KW - Crystal structure

KW - Graphene nanoparticles

KW - Kunitz–type trypsin inhibitor

KW - Zinc oxide nanoparticles

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U2 - 10.1016/j.csbj.2024.07.023

DO - 10.1016/j.csbj.2024.07.023

M3 - Journal article

AN - SCOPUS:85201013190

SN - 2001-0370

VL - 23

SP - 3132

EP - 3142

JO - Computational and Structural Biotechnology Journal

JF - Computational and Structural Biotechnology Journal

ER -

Crystal structure of Kunitz-type trypsin inhibitor: Entomotoxic effect of native and encapsulated protein targeting gut trypsin of Tribolium castaneum Herbst

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