Metals in Medicine Network NSW Regional Meeting Structural Biology Mitchell Guss School of Molecular & Microbial Biosciences University of Sydney 19 Jan.

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Presentation transcript:

Metals in Medicine Network NSW Regional Meeting Structural Biology Mitchell Guss School of Molecular & Microbial Biosciences University of Sydney 19 Jan 2004

Structural Biology in Australia Limited to X-ray and NMR of macromolecules (may not be accurate) NSW (X-ray:2*, NMR:3(4)) ACT (X-ray:1, NMR:1) VIC (X-ray:5, NMR:2) QLD (X-ray:3(4), NMR:2) SA (X-ray:1, NMR:1) WA (X-ray:1, NMR:1) *sites not PI’s

Sydney: Historical Outline : Metal amino-acid and metal peptide complexes (Chemistry: Freeman) : Metalloprotein structures (Chemistry: Freeman, Guss & Wright) 1994-current: Metalloprotein structures (MMB: Freeman, Guss, Mackay, Maher, Matthews)

Selected Highlights Blue copper proteins –Plastocyanin (1 st new protein structure solved in Australia) –Cucumber Basic Protein (1 st metalloprotein structure solved by the multiple wavelength anomalous scattering method) –Auracyanin Iron proteins –Legume hemoglobin –Rubredoxin (mutants and different metals) –Purple acid phosphatase Copper enzymes –Amine oxidases (pea seedling, Arthrobacter globiformis, Pichia pastoris) Others –Aminopeptidase P (manganese) –Prolidase (cobalt, zinc) –Selenate reductase (selenium) –Dihydroorotase (zinc) Transcription Factors (zinc) Oligonucleotides (cobalt)

Principal Techniques Nuclear magnetic resonance spectroscopy (Mackay, Matthews, Kuchel) –400 MHz, 600 MHz (MMB, cryoprobe 2004/5) –800 MHz (ANU, 2004) X-ray crystallography (Collyer, Freeman, Guss, Maher) –Rotating anode generator, Marresearch image plate detectors (MMB) –Synchrotron (SSRL, APS)

Facilities for Structure Characterisation - MMB Ultracentrifugation Plasmon resonance CD spectroscopy Microcalorimetry Mass spectrometry (Electrospray and Maldi) Fluorescence spectroscopy Dynamic and multi-angle light scattering

Additional Facilities - MMB Protein expression and purification (Fermentation for bacterial and mammalian* cells) Capillary electrophoresis FPLC and HPLC Proteomics (2D gels, automated spot picking, mass spectrometry) Animal house PC2 & PC3 labs *only bacterial expression to date

Computing - MMB X-ray & NMR groups share a mixed LINUX/IRIX cluster 6 Intel-based LINUX servers providing file and compute resources (100 GBytes user and 200 GBytes of scratch storage; 10 cpus) 6 LINUX workstations; 7 IRIX (SGI- O2) workstations for graphics applications

Collaboration or Service? Protein structure determination is becoming increasing “routine”: –Still requires major commitment of personnel resources –Not a service –Close collaboration Growing focus on the biology of systems –Use of whatever techniques required to solve the problem Lack of technical support staff to enable routine (casual) use of instrumentation