In vitro cross-species membrane insertion of Neisserial Nalp by Campylobacter jejuni-BamA (360G-Wellcome-202374_Z_16_Z)

£2,000

Outer membrane proteins (OMPs) play a fundamental role in physiological functions, multidrug resistance and virulence in Gram-negative bacteria making them essential for cell viability in a number of pathogens. Biogenesis of such beta-barrel proteins is facilitated by BamA, the main hub of the Barrel Assembly Machinery (BAM) complex. While structural characterisation of the MAS complex is yielding first results on the E. coli system (Backelar et al 2016), mechanistic details remain notoriously difficult to obtain. We propose to investigate the role of BamA from Campylobacter jejuni, an important pathogen and food spoilage organism, as a facilitator of membrane insertion of an autotransporter translocator domain from Neisseria meningitidis, NalP. NalP is a beta-barrel, which can insert at low levels spontaneously into lipid membranes in a membrane composition-dependent way. We propose to use our homology model of C. jejuni, annealed by molecular dynamics simulations (MD), to perform in silico CHARMM docking of NalP. We will investigate experimentally the degree of NalP insertion into model lipid membranes without (liposomes) or with BamA (proteoliposomes). We will use gradient separation and SDS-PAGE to quantify insertion, as well as solid state and DNP NMR to confirm insertion and to investigate specific interactions.

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Grant Details

Amount Awarded 2000
Applicant Surname Sciences
Approval Committee Internal Decision Panel for C&S
Award Date 2016-04-01T00:00:00+00:00
Financial Year 2015/16
Grant Programme: Title Vacation Scholarships
Internal ID 202374/Z/16/Z
Lead Applicant Mr School Sciences
Partnership Value 2000
Planned Dates: End Date 2016-08-12T00:00:00+00:00
Planned Dates: Start Date 2016-06-13T00:00:00+00:00
Recipient Org: Country United Kingdom
Region East Midlands