Biophysics

IRIC’s Biophysics core facility provides services in cutting-edge solution state NMR spectroscopy with a specific emphasis on experiments investigating protein structure, protein-ligand interactions and routine small molecule analysis.

The platform has two Varian NMR spectrometers: a 600 MHz INOVA equipped with an HCN coldprobe mainly dedicated to large molecule (protein/nucleic acid) analysis and a 400 MHz MR system with an autosampler routinely used for small molecule applications.

 

 

Services offered

We provide services in acquisition of NMR experiments and assistance in analysis of spectra. We also offer users advice and training in:

  • Expression, stable isotope incorporation and purification of proteins
  • Strategies for structure determination of proteins and small molecules and investigations of protein-ligand complexes
  • Processing and interpretation of NMR spectra
  • Softwares to generate 3D structures from NMR data
  • Automated acquisition of spectra for small molecules

Advice and implementation of NMR experiments for the assignment and structure determination of proteins, theses include (but are not restricted to):

  • Spectra for backbone assignment; HNCACB, HNCA, CBCA(CO)NH, HNCOCA, HNCO, etc.
  • Spectra for sidechain assignment; C(CO)NH-TOCSY, HC(CO)NH-TOCSY, HCCH-TOCSY etc
  • Spectra for production of structural restraints; 2D, 3D, 4D 15N and 13C-edited NOESY’s, HNHA, HNHB 
  • Implementation of state-of-the-art methods for expediating NMR acquisiton including non-linear acquisition and FAST and SO-FAST methods
  • Determination of residual dipolar couplings in aligned media; IPAP-HSQC
  • Spectra for investigating small molecule binding to proteins; 1D and 2D STD experiments, 2D TCS experiments, 1D T1rho experiments, 2D 1H-15N HSQC titration experiments. 3D select filter-edit experiments
  • Advice on expression, uniform labelling and purification of proteins 
  • Advice and help on processing and interpretation of NMR spectra
  • Access to in-house scripts to aid analysis of NMR data

We also provide training for users to use the automated 400 MHz NMR system for routine spectra of small molecules including:

  • 1D spectra of nuclei ranging from 15N to 31P, including 1H
  • 2D spectra including COSY, NOESY, ROESY, HSQC, HMQC, HMBC, H2BC experiments

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