Projects per year
Abstract
The membrane protein DsbD is a reductase that acts as an electron hub, translocating reducing equivalents from cytoplasmic thioredoxin to a number of periplasmic substrates involved in oxidative protein folding, cytochrome c maturation and oxidative stress defence. DsbD is a multi-domain protein consisting of a transmembrane domain (t-DsbD) flanked by two periplasmic domains (n-DsbD and c-DsbD). Previous studies have shown that DsbD is required for the survival of the obligate human pathogen Neisseria meningitidis. To help understand the structural and functional aspects of N. meningitidis DsbD, the two periplasmic domains which are required for electron transfer are being studied. Here, the expression, purification and biophysical properties of n-NmDsbD and c-NmDsbD are described. The crystallization and crystallographic analysis of n-NmDsbD and c-NmDsbD are also described in both redox states, which differ only in the presence or absence of a disulfide bond but which crystallized in completely different conditions. Crystals of n-NmDsbDOx, n-NmDsbDRed, c-NmDsbDOx and c-NmDsbDRed diffracted to 2.3, 1.6, 2.3 and 1.7 resolution and belonged to space groups P213, P321, P41 and P1211, respectively.The cloning, purification, biophysical characterization, crystallization and X-ray diffraction analysis of the two periplasmic domains of Neisseria meningitidis DsbD, a disulfide reductase that is essential for the viability of this human pathogen, are reported.
Original language | English |
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Pages (from-to) | 31-38 |
Number of pages | 8 |
Journal | Acta Crystallographica Section F: Structural Biology Communications |
Volume | 74 |
Issue number | 1 |
DOIs | |
Publication status | Published - 1 Jan 2018 |
Keywords
- disulfide catalysis
- DsbD
- membrane proteins
- Neisseria meningitidis
Projects
- 2 Finished
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DsbA inhibitors: from hits to leads
Martin, J. L., Scanlon, M., Heras, B., Scammells, P. & Torsika, M.
1/01/16 → 31/12/18
Project: Research
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Characterisation of antigenic variation of Neisserial cell surface adhesins, and thir role in infection
Davies, J. & Kahler, C.
National Health and Medical Research Council (NHMRC) (Australia)
5/01/09 → 31/12/12
Project: Research
Equipment
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Australian Synchrotron
Office of the Vice-Provost (Research and Research Infrastructure)Facility/equipment: Facility