Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.523186
Title: Protein mis-folding and human disease
Author: Pal, Mohinder
Awarding Body: University of Southampton
Current Institution: University of Southampton
Date of Award: 2010
Availability of Full Text:
Access through EThOS:
Access through Institution:
Abstract:
Serum Amyloid P Component (SAP), a putative molecular chaperone, is a homopentameric plasma protein of 25kDa subunits. It binds to the amyloid fibrils of misfolded proteins, which cause amyloidosis in humans. SAP not only stabilizes amyloid fibrils but also protects them from proteolytic and cell mediated degradation. SAP has been co-crystallized with three different aminoalkyl phosphonates that bind at the amyloid recognition site of SAP, and the X-ray crystal structures were determined at atomic resolution. A secondary aim of this work was to understand the enhanced amyloidogenic potential of L55P and V30M transthyretin (TTR) protein. TTR misfolding has been implicated in number of human diseases such as senile systemic amyloidosis, familial amyloid polyneuropathy and familial amyloid cardiopathy. TTR protein is a thyroxine binding protein (14kDa) existing as a tetramer in vivo. L55P and V30M mutant TTR are the most aggressive and most common mutants, respectively, in causing FAP. L55P and V30M mutant TTR protein were expressed in E.coli and purified using anion-exchange chromatography and gel filtration. L55P and V30M TTR were co-crystallised with MDS84, a compound that has been demonstrated to stabilize the tetramer in vitro. The X-ray structures of L55P and V30M, TTR mutant proteins have been determined at 1.5Å and 2.1Å resolution. In addition, research was carried out on the bacterial protein Burkholderia invasion protein D (BipD). BipD (33kDa) belongs to type III secretion system of Burkholderia pseudomallei. It creates a pore in the host cell membrane to help the B. pseudomallei invasion. This bacterial infection causes melioidosis disease in humans. To determine the ligand recognition site of BipD, its high-resolution crystal structure has been determined at 1.5Å. This high resolution BipD structure is more complete than its previously solved structures and is in the new space group C2. The BipD structure presented in this research may help to design potential chemical inhibitors of BipD to prevent bacterial invasion into human body. The molecular chaperones play an important role in the protein refolding and assembly. ATJ11 is of 14kDa protein belongs to DnaJ co-chaperone family. The protein has been expressed in E.coli and purified by affinity and cation exchange chromatography. The biophysical study of ATJ11 (CD spectrum) has been conducted showing its predominant α-helical structure and attempts were made to crystallise it but no protein crystals have been obtained.
Supervisor: Wood, Steve P. ; Coker, A. R. Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.523186  DOI: Not available
Keywords: RB Pathology ; QH301 Biology
Share: