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Developing an innovative approach to identify novel talin-binding ligands in the brain

Albanese, Valentina (2023) Developing an innovative approach to identify novel talin-binding ligands in the brain. Master of Research (MRes) thesis, University of Kent. (doi:10.22024/UniKent/01.02.101809) (Access to this publication is currently restricted. You may be able to access a copy if URLs are provided) (KAR id:101809)

Language: English

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Cell adhesion, a process underlying the maintenance and development of multicellularity, is fundamentally dependent on the activity of talin as a mechanosensitive signalling nexus. Talin couples integrin-mediated cell-to-matrix adhesions to the actin cytoskeleton, facilitating the formation of integrin adhesion complexes, which act as sophisticated information processing centres. Variation of talin rod domain conformation, following adhesion, promotes different talin-mediated recruitment and signalling outputs, enabling the modification of a cell's internal programming. The recently published MeshCODE theory describes how information in the brain may be stored binarily via the mechanicalistic alteration of talin rod binary switches, highly enriched at every synapse. To validate this theory and discover a connection between neuronal activity and talin-mediated mechanical signalling, an innovative method, termed the talin 'fishing' rod experiment, was developed to identify novel talin-binding ligands in murine neuronal extract. Many proteins were identified as promising neuronal talin-binding ligands, including dynamin-1, synaptojanin-1, and myosin-Va, all of which exhibit particular significance to many aspects of synaptic transmission, particularly in the regulation of synaptic vesicle endo- and exocytosis. A bioinformatics pipeline and subsequent scoring system was also developed to accompany this experimental approach, enabling the identification of probable talin-binding sites in known synaptic vesicle proteins through I/LD motif evaluation. Whilst only weak binding was demonstrated between several talin rod domains and the proteins selected through bioinformatics analysis, it implies that both the bioinformatics pipeline and scoring system is effective in identifying promising talin-binding I/LD motifs. The identification of adhesome proteins, prior to this project, was restricted to cell lines and fibroblasts, limiting our knowledge of talin-binding ligands outside of these cell types. However, the talin 'fishing' rod experiment, developed in this project, can be utilised to help fill this current knowledge gap. Validation of the promising interactors, identified in this study, will support the concepts presented in the MeshCODE theory, potentially enabling the scientific community to achieve understanding regarding the enigma of memory storage at long last.

Item Type: Thesis (Master of Research (MRes))
Thesis advisor: Goult, Benjamin T
DOI/Identification number: 10.22024/UniKent/01.02.101809
Subjects: Q Science
Divisions: Divisions > Division of Natural Sciences > Biosciences
Depositing User: System Moodle
Date Deposited: 23 Jun 2023 11:47 UTC
Last Modified: 27 Jun 2023 07:35 UTC
Resource URI: (The current URI for this page, for reference purposes)

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