Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.677143
Title: Unearthing the molecular mechanisms that govern L-selectin-dependent adhesion and migration
Author: Newe, Abigail Lucy
ISNI:       0000 0004 5368 3829
Awarding Body: King's College London
Current Institution: King's College London (University of London)
Date of Award: 2015
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Abstract:
L-selectin has been well characterised as a cell adhesion molecule, which plays a role in the recruitment of leukocytes to sites of inflammation and is responsible for the recirculation of lymphocytes to secondary lymphoid organs. Recent evidence has shown that L-selectin also acts as a signalling molecule to activate pathways and regulate the inflammatory response. The cytosolic tail of L-selectin plays a crucial role in regulating its activity through its interaction with binding partners, such as calmodulin (CaM) and the ERM protein family. However, little is known about how the interaction between L-selectin and its binding partners is regulated. The aim of this multidisciplinary PhD project is to use biophysical and cell biological methods to address the role of the interaction between L-selectin and its binding partners during leukocyte recruitment. To this end, the interaction between CaM and the L-selectin cytosolic tail was assessed using isothermal titration calorimetry (ITC) and nuclear magnetic resonance (NMR) spectroscopy. Analysis revealed that phosphorylation of serine residues within the cytosolic tail of L-selectin did not affect CaM binding. To enable the observation of the interaction between L-selectin and CaM whilst leukocytes are undergoing transendothelial migration (TEM), the THP-1 monocytic cell line was engineered to stably express L-selectin-GFP and CaM-RFP so their interaction could be monitored at different stages of TEM. The data showed that phosphorylation of serine 364 in the L-selectin tail is important for regulating CaM interaction. Discrepancies were identified between the biophysical and cell biological results, implying the leukocyte plasma membrane may play a vital role in regulating the interaction between L-selectin and CaM. This highlights the importance of studying transmembrane proteins in the correct context.
Supervisor: Ivetic, Aleksandar ; Conte, Maria Rosaria Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.677143  DOI: Not available
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