jonathan worrall mcb away day 25.09.13. an appraisal of metals used in proteins systematic survey of...
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Jonathan WorrallMCB away day
25.09.13
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An appraisal of metals used in proteins
Systematic survey of 1371 different enzymes with a known 3D structure
47% required metals with 41% requiring metals at their catalytic sites
JBIC (2008) 13, 1205-1218Nature (2009) 460, 823-830
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Research Interests
Microbial biotechnologyApoptosis
Cytochrome c/phospholipid complexes Streptomyces lividans
Cu
Worrall & Vijgenboom 2010, Nat. Prod. Reps. 5, 742.
Cuproproteins and enzymes must havea role in triggering development switches
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R580 hours
R5 10 μM Cu(II)80 hours
S. lividans
R5140 hours
ΔSLI4214
ΔSLI4213
ΔSLI4212
ΔSLI3188
DCuproproteins/enzymes have morphological phenotypes
Blundell et al. 2013, Open Biology 3, 120163
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Ty act / biomass
-500
0
500
1000
1500
2000
2500
3000
3500
4000
0 20 40 60
Time (hour)Ty
act
ivity
(uni
ts/m
in,g
,lit)
1326 pIJ703
2837::Apra pIJ703
DSLI3188
S. lividans
Morphological Engineering
Reengineering of Streptomyces strains as hosts for synthetic biology approaches to create sustainable industrial production platforms
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Techniques used
• Molecular biology• Recombinant protein over-expression • Protein chromatography• UV-visible, circular dichroism (CD) spectroscopies • Electron paramagnetic resonance (EPR) spectroscopy• Nuclear magnetic resonance (NMR) spectroscopy (UEA)• X-ray crystallography• Isothermal titration calorimetry (ITC)• Stopped-flow kinetics• Electrophoretic mobility shift assays (EMSA)
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Collaborations
UEA (Cytochrome c)Professor Geoff Moore (NMR)
Professor Julea Butt (Electrochemistry)
Molecular Biotechnology, Leiden, NLDr Erik Vijgenboom
EssexDr Mike Hough (X-ray)
Dr Dimitri Svistunenko (EPR)Professor Mike Wilson (SF-kinetics)