Shotgun Proteomic-Based Approach with a Q-Exactive Hybrid Quadrupole-Orbitrap High-Resolution Mass Spectrometer for Protein Adductomics on a 3D Human Brain Tumor Neurospheroid Culture Model: The Identification of Adduct Formation in Calmodulin-Dependent Protein Kinase-2 and Annexin-A1 Induced by Pesticide Mixture.

Autor: Louati K; Faculty of Pharmacy, Laboratory of Pharmacology, Analytics & Galenic Drug Development-LR12ES09, University of Monastir, Road Avicenne, Monastir 5000, Tunisia., Maalej A; Laboratory of Environmental Bioprocesses, Centre of Biotechnology of Sfax, Road of Sidi-Mansour, P.O. Box 1177, Sfax 3018, Tunisia., Kolsi F; Department of Neurosurgery, Habib Bourguiba University Hospital, Road El Ain km 1.5, Avenue of Ferdaous, Sfax 3089, Tunisia.; Faculty of Medicine, Avenue of Majida Boulila, University of sfax, Sfax 3029, Tunisia., Kallel R; Laboratory of Pathological Anatomy and Cytology, Habib Bourguiba University Hospital, Road El Ain km 1.5, Avenue of Ferdaous, Sfax 3089, Tunisia.; Faculty of Medicine, Avenue of Majida Boulila, University of sfax, Sfax 3029, Tunisia., Gdoura Y; Department of Neurosurgery, Habib Bourguiba University Hospital, Road El Ain km 1.5, Avenue of Ferdaous, Sfax 3089, Tunisia.; Faculty of Medicine, Avenue of Majida Boulila, University of sfax, Sfax 3029, Tunisia., Borni M; Department of Neurosurgery, Habib Bourguiba University Hospital, Road El Ain km 1.5, Avenue of Ferdaous, Sfax 3089, Tunisia.; Faculty of Medicine, Avenue of Majida Boulila, University of sfax, Sfax 3029, Tunisia., Hakim LS; Laboratory of Pathological Anatomy and Cytology, Habib Bourguiba University Hospital, Road El Ain km 1.5, Avenue of Ferdaous, Sfax 3089, Tunisia., Zribi R; Higher Institute of Applied Studies to Humanities of Tunis (ISEAHT), University of Tunis, 11 Road of Jebel Lakdhar, Tunis 1005, Tunisia., Choura S; Laboratory of Environmental Bioprocesses, Centre of Biotechnology of Sfax, Road of Sidi-Mansour, P.O. Box 1177, Sfax 3018, Tunisia., Sayadi S; Biotechnology Program, Center for Sustainable Development, College of Arts and Sciences, Qatar University, Doha 2713, Qatar., Chamkha M; Laboratory of Environmental Bioprocesses, Centre of Biotechnology of Sfax, Road of Sidi-Mansour, P.O. Box 1177, Sfax 3018, Tunisia., Mnif B; Department of Bacteriology, Habib Bourguiba University Hospital, Road El Ain km 1.5, Avenue of Ferdaous, Sfax 3089, Tunisia.; Faculty of Medicine, Avenue of Majida Boulila, University of sfax, Sfax 3029, Tunisia., Khemakhem Z; Legal Medicine Department, Habib Bourguiba University Hospital, Road El Ain km 1.5, Avenue of Ferdaous, Sfax 3089, Tunisia.; Faculty of Medicine, Avenue of Majida Boulila, University of sfax, Sfax 3029, Tunisia., Boudawara TS; Laboratory of Pathological Anatomy and Cytology, Habib Bourguiba University Hospital, Road El Ain km 1.5, Avenue of Ferdaous, Sfax 3089, Tunisia.; Faculty of Medicine, Avenue of Majida Boulila, University of sfax, Sfax 3029, Tunisia., Boudawara MZ; Department of Neurosurgery, Habib Bourguiba University Hospital, Road El Ain km 1.5, Avenue of Ferdaous, Sfax 3089, Tunisia.; Faculty of Medicine, Avenue of Majida Boulila, University of sfax, Sfax 3029, Tunisia., Safta F; Faculty of Pharmacy, Laboratory of Pharmacology, Analytics & Galenic Drug Development-LR12ES09, University of Monastir, Road Avicenne, Monastir 5000, Tunisia.
Jazyk: angličtina
Zdroj: Journal of proteome research [J Proteome Res] 2023 Dec 01; Vol. 22 (12), pp. 3811-3832. Date of Electronic Publication: 2023 Oct 31.
DOI: 10.1021/acs.jproteome.3c00484
Abstrakt: Pesticides are increasingly used in combinations in crop protection, resulting in enhanced toxicities for various organisms. Although protein adductomics is challenging, it remains a powerful bioanalytical tool to check environmental exposure and characterize xenobiotic adducts as putative toxicity biomarkers with high accuracy, facilitated by recent advances in proteomic methodologies and a mass spectrometry high-throughput technique. The present study aims to predict the potential neurotoxicity effect of imidacloprid and λ-cyhalothrin insecticides on human neural cells. Our protocol consisted first of 3D in vitro developing neurospheroids derived from human brain tumors and then treatment by pesticide mixture. Furthermore, we adopted a bottom-up proteomic-based approach using nanoflow ultraperformance liquid chromatography coupled with a high-resolution mass spectrometer for protein-adduct analysis with prediction of altered sites. Two proteins were selected, namely, calcium-calmodulin-dependent protein kinase-II (CaMK2) and annexin-A1 (ANXA1), as key targets endowed with primordial roles. De novo sequencing revealed several adduct formations in the active site of 82-ANXA1 and 228-CaMK2 as a result of neurotoxicity, predicted by the added mass shifts for the structure of electrophilic precursors. To the best of our knowledge, our study is the first to adopt a proteomic-based approach to investigate in depth pesticide molecular interactions and their potential to adduct proteins which play a crucial role in the neurotoxicity mechanism.
Databáze: MEDLINE
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