Use this URL to cite or link to this record in EThOS:
Title: Development of nuclear magnetic resonance methods for determination of membrane protein structure
Author: Patel, Dharmesh K.
ISNI:       0000 0004 2748 8229
Awarding Body: University of Warwick
Current Institution: University of Warwick
Date of Award: 2012
Availability of Full Text:
Access from EThOS:
Access from Institution:
Membrane proteins represent over a third of all proteins encoded for by the human genome and play a vital role in the functionality of the cell, by controlling a vast number of cellular processes. With over half of pharmacological drugs targeting membrane proteins, their importance is not to be under estimated. Yet the number of three-dimensional membrane protein structures reported to date falls well short of that of their water soluble counterparts. This discrepancy can directly be attributed to the difficulties involved in studying membrane protein structure due to their hydrophobic nature, resulting in a number of challenges in the production and purification of protein, whilst requiring the use of a suitable membrane mimetic upon extraction from their native membrane. Solid state NMR (ssNMR) as a technique for studying membrane protein structure is well placed in being able to obtain structural information for membrane proteins in “native-like” lamellar bilayer environments but there are challenges involved in preparing suitable samples for analysis. As there is no “one suit fits all” method for preparing membrane protein samples for ssNMR analysis, conditions that result in fully reconstituted protein, that also allow for high resolution structural analysis have to be trialled. This study presents work on sample preparation methods for the reconstitution of the small alpha helical transmembrane (TM) proteins, using the well characterised TM protein Glycophorin A (GpA) as a model peptide. Established biophysical and NMR techniques were used to characterise DMPC lipid embedded peptides prepared using two reconstitution techniques. The limited site specific labelling at key positions of the GpA homodimer was used to evaluate the feasibility of using similar sample preparation and labelling schemes when applied to that of the Bovine Papillomavirus E5 (BPV E5) TM protein, for which no solved three-dimensional structure exists. Characterisation of the DMPC membranes into which membrane proteins where reconstituted was also conducted. To compliment ssNMR analysis of BPV E5, preliminary work on the use of fast tumbling isotropic bicelles to study membrane protein structure by solution NMR is also presented.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available
Keywords: QD Chemistry ; QP Physiology