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Title: Structural studies on immunogenic proteins of Burkholderia pseudomallei
Author: Pramanpol, Nuttawan
ISNI:       0000 0004 2736 3725
Awarding Body: University of Sheffield
Current Institution: University of Sheffield
Date of Award: 2013
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Burkholderia pseudomallei is a Gram negative bacterial pathogen and the causative agent of melioidosis, which is the third most frequent cause of death from infectious disease in Northeastern Thailand and also highly endemic in Southeast Asia and Northern Australia. Problems of melioidosis management include misdiagnosis and the increasing resistance of the bacteria to the current anti-infective treatment, resulting in high mortality and having a global impact as a potential bioweapon. Little is known about the ambiguous pathogenesis and virulence mechanisms of B. pseudomallei infection. One way to gain understanding is through the investigation of the structure/function relationships of potential target proteins, as a starting point for a rational drug discovery programme. Structural studies on the target proteins of B. pseudomallei can shed light on their biological functions at the molecular level. This thesis presents the structural studies on immunogenic proteins of B. pseudomallei as part of a targeted structural genomics programme on B. pseudomallei. Seven target genes were selected from lists of putative essential genes and also identified B. pseudomallei proteins that elicit an immuno cross reactivity to the sera of infected humans, which may be involved in virulence and/or pathogenicity of B. pseudomallei. Of those, BPSL0606, an uncharacterised protein, was cloned, over-expressed, purified, and crystallised. The first crystal structure of BPSL0606 was determined and revealed that it is structurally similar to the GCN5-related N-acetyltransferase (GNAT) superfamily. Dimer formation in the crystal packing agreed with gel filtration analysis and the interface of BPSL0606 was similar to many GNAT members. Structure-based alignment of BPSL0606 against known structures of GNATs showed that BPSL0606 has conserved residues that are involved in the binding of acetyl coenzyme A in members of GNATs, suggesting that BPSL0606 may bind the acetyl coenzyme A. Although co-crystallization of BPSL0606 and acetyl coenzyme A and ligand observation by nuclear magnetic resonance suggested that BPSL0606 did not bind to the acetyl coenzyme A. Analysis of the aminoglycoside binding site indicated that BPSL0606 could possibly bind to this substrate. The BPSL0606 structure with acetyl coenzyme A and kanamycin was modelled, proposing a possible clue related to the antibiotic resistance. The findings in this PhD thesis provide the molecular and structural information of B. pseudomallei proteins, which could contribute to further investigations.
Supervisor: Baker, Patrick Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available