the analytical laboratory school of biological sciences dr bowater prof. richardson interactions of...

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The analytical laboratory School of Biological Sciences Dr Bowater Prof. Richardson Interactions of proteins with DNA The bacterial nitrogen cycle & anaerobic metabolism of bacterial pathogens Dr Brearley Dr Watmough Plant metabolic biochemistry Enzymology of Nitric oxide generation/metabolism Oxidases and peroxidases in aerobic metabolism Dr Clarke Biophysical/spectroscopic analysis of protein-protein interactions Dr Hemmings X-ray structural and mechanistic studies of enzymes Catalytic properties of metalloproteins involved in bacterial respiration which enable bacteria to exploit host cell environments

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Page 1: The analytical laboratory School of Biological Sciences Dr Bowater Prof. Richardson Interactions of proteins with DNA The bacterial nitrogen cycle & anaerobic

The analytical laboratorySchool of Biological Sciences

Dr Bowater Prof. RichardsonInteractions of proteins with DNA The bacterial nitrogen cycle & anaerobic metabolism of bacterial pathogens

Dr Brearley Dr WatmoughPlant metabolic biochemistry Enzymology of Nitric oxide generation/metabolism Oxidases and peroxidases in aerobic metabolism Dr ClarkeBiophysical/spectroscopic analysis of protein-protein interactions

Dr HemmingsX-ray structural and mechanistic studies of enzymes

Catalytic properties of metalloproteins involved in bacterial respiration which enable bacteria

to exploit host cell environments in symbiosis and pathogenesis

Page 2: The analytical laboratory School of Biological Sciences Dr Bowater Prof. Richardson Interactions of proteins with DNA The bacterial nitrogen cycle & anaerobic

The analytical laboratorySchool of Biological Sciences

Facilities & expertise

Protein expression.

Protein purification

Protein characterization

Chromatographic analysis/purification of small molecules

Page 3: The analytical laboratory School of Biological Sciences Dr Bowater Prof. Richardson Interactions of proteins with DNA The bacterial nitrogen cycle & anaerobic

The analytical laboratorySchool of Biological Sciences

Protein purification

Under aerobic and anaerobic conditionsfrom Category 2 microorganismsLicense/authority from Defra, HSE

Protein purification on analytical, pilot and larger scales

Page 4: The analytical laboratory School of Biological Sciences Dr Bowater Prof. Richardson Interactions of proteins with DNA The bacterial nitrogen cycle & anaerobic

Protein structure determinationX-ray crystallography with access to beam time on synchrotonsRobotic crystallization trials

Protein characterization - capabilities

Structure-informed protein characterization and rationale design is available

Page 5: The analytical laboratory School of Biological Sciences Dr Bowater Prof. Richardson Interactions of proteins with DNA The bacterial nitrogen cycle & anaerobic

Protein structure prediction and modellingEnzymology, especially of metalloproteinsVariety of spectroscopic techniquesTime-resolved spectroscopy of metalloproteinsFast reaction kinetics

Analytical ultra centrifugationHydrodynamic properties of proteins

Protein characterization - capabilities

In absence of crystal structures, protein structures are modelled

Page 6: The analytical laboratory School of Biological Sciences Dr Bowater Prof. Richardson Interactions of proteins with DNA The bacterial nitrogen cycle & anaerobic

Isothermal calorimetryto study protein-ligand interactions

True thermodynamic parameters for any reaction are obtained by ITC

0.0 0.5 1.0 1.5 2.0 2.5

-10

-8

-6

-4

-2

0

0 10 20

-0.4

-0.2

0.0

0.3 mg/ml (C12A)Mur + 1 mMMn(II)Time (min)

µca

l/sec

Molar Ratio

kcal

/mol

e of

inje

ctan

t

Page 7: The analytical laboratory School of Biological Sciences Dr Bowater Prof. Richardson Interactions of proteins with DNA The bacterial nitrogen cycle & anaerobic

Metabolic and small molecule analysis

Gas Chromatography with liquid and headspace samplingElectron Impact GC-Mass Spectrometry Flame Ionization Detection Electron Capture Detection

Fermentation products, gases, natural oils are analyzed by GC

Page 8: The analytical laboratory School of Biological Sciences Dr Bowater Prof. Richardson Interactions of proteins with DNA The bacterial nitrogen cycle & anaerobic

Metabolic and small molecule analysis

Tracer analysisStable isotopes and radioisotopesLicensed by Env. Agency

High Pressure Liquid Chromatograhy

on analytical and semi-prep scale

UV, DAD, suppressed ion conductivity, fluorescence &

radiochemical detection

HPLC-Mass Spectrometry

HPLC method development is available