Ratkeviciute, Giedre 
ORCID: 0000-0003-1682-1455
(2024).
Structural investigation of putative lipid transport systems in Bdellovibrio bacteriovorus HD100.
University of Birmingham.
Ph.D.
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Ratkeviciute2024PhD.pdf
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Abstract
Mammalian Cell Entry (MCE) domain-containing proteins have been implicated in glycerophospholipid transport, a key component of the gram-negative OM. The model organism Escherichia coli contains three MCE proteins residing within unique operons; MlaD, PqiB and LetB, each presenting potential novel targets for innovative antibacterial agents. Characterising these proteins and their associated complexes within multiple organisms may provide additional insight into these putative lipid transport systems.
Here, we focus on MCE proteins within Bdellovibrio bacteriovorus HD100, a predatory gram- negative bacterium, that alternates between free-living growth and predation upon other gram-negative bacteria. Thus far, we investigated Bd1152, a two-MCE domain containing protein residing at the centre of a tripartite operon.
The inherent structural plasticity of Bd1152 posed a challenge in obtaining its high-resolution structure. Nevertheless, we present a crystal structure of Bd1152 as well as a cryo-EM structure of the Bd1152 top MCE ring at 2.6 Å and 3.4 Å resolution, respectively. Both structures reveal hexameric assemblies (MCE rings) with a central channel that has been implicated in phospholipid transport. Furthermore, we utilised the novel Zernike3D and cryoDRGN algorithms to compute inherent continuous flexibility, subsequently revealing that Bd1152 possesses high structural plasticity, potentially providing insight into the mechanism of lipid transport.
| Type of Work: | Thesis (Doctorates > Ph.D.) | |||||||||
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| Award Type: | Doctorates > Ph.D. | |||||||||
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| Licence: | All rights reserved | |||||||||
| College/Faculty: | Colleges (2008 onwards) > College of Life & Environmental Sciences | |||||||||
| School or Department: | School of Biosciences | |||||||||
| Funders: | Biotechnology and Biological Sciences Research Council, Other | |||||||||
| Other Funders: | Midlands Integrative Biosciences Training Programme studentship (grant no. BB/M01116jdjhashfirX/1), BBSRC and University of Birmingham, BBSRC Research grant (grant no. BB/S017283/1), Instruct-ERIC (grant no. PID2350 and PID23275) | |||||||||
| Subjects: | Q Science > Q Science (General) | |||||||||
| URI: | http://etheses.bham.ac.uk/id/eprint/14429 |
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