The biophysics platform offers state-of-the-art instruments to investigate the physical and structural properties of biologically important molecules; including potential therapeutics (small molecules) and drug targets (protein, RNA/DNA) at the structural level, in addition to characterization of biologically relevant interactions. Screening services to identify potential therapeutic molecular using NMR-based techniques with small molecule fragment libraries are offered.
Services offered:
- Analysis of samples using NMR and analysis of spectra
- Access to in-house scripts to aid the analysis of NMR data
- User advice and training in:
- Incorporation of stable isotopes and purification of proteins
- Developing strategies for structure determination of proteins and small molecules, and investigation of protein-ligand complexes
- Processing and interpretation of NMR spectra
- Use of software to generate 3D structures from NMR study data
- Automated acquisition of spectra for small molecules
- 1D spectra of nuclei ranging from 15N to 31P, including 1H
- 2D spectra, including COSY, NOESY, ROESY, HSQC, HMQC, HMBC, H2BC experiments
- Implementation of NMR experiences for the assignment and structure determination of proteins, including (but 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, HNHA, HNHB
- Implementation of innovative methods for expediating NMR experiments including non-linear acquisitions and SO-FAST methods
- Determination of dipolar couplings; IPAP-HSQC
- Access to 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
Equipment
600 MHz NMR spectrometer
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- Magnet and Console
- Bruker Avance III HD 600 MHz. 4 channel system capable of 2H decoupling while simultaneously acquiring three other nuclei (1H, 13C, 15N typically, also 31P).
- Probes
- 5mm QCIP with Z gradients. Cryoprobe optimized for simultaneous observation of 1H(19F), 13C, 15N and 31P. Compatible with 3mm tubes. This is installed 99% of the time
- 1.7mm QCI with Z gradients. Suitable for highly
- Sundries
- Bruker Sample Jet
- Accommodates up to 480 samples using five 96-well-plate-format racks (5mm, 3mm, 1.7mm tubes). Integrated with automation software allowing high throughput protocols. Temperature controlled: ideal for fragile biological systems.
- Applications
- High sensitivity with a cryo-probe makes this system ideal to study complicated biological systems such as proteins, DNA/RNA and their interactions. Additionally small molecules with limited amounts or limited solubility can be analyzed for both 1H and 13C nuclei. Typically used for multidimensional, triple resonance experiments for assignments, structure determination and dynamics studies of proteins, nucleic acids and their interactions. The samplejet with automated software (ICONNMR) permits for high throughput analysis of samples allowing application of NMR-based fragment screening.
- Typical expts offered
- Triple resonance experiments (2H decouplng if needed) for routine protein NMR assignment of folded/unfolded proteins
- FAST HSQC experiments (with AI optimized pulses) for 15N/13C HSQC experiments
- 15N relaxation experiments to investigate dynamics in proteins
- Ligand and protein based fragment screens (Library – Maybridge available)
- Quantitation (accurate concentration measurement) of small molecules
- Analysis
- Advice for protein assignment and structure determination; including using automated softwares (ARTINA)
- Relaxation data analysis; T1 T2 NOE CPMG, CEST, relaxation dispersion
- Determination of association constants (Kd)
- PCA analysis of spectra
- Identification of binders from NMR based fragment screens
- Small molecule characterization using NOAH techniques
2. 400 MHz NMR spectrometer
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- Magnet and Console
- Bruker Avance NEO 400 MHz. 2 channel system suitable for double resonance experiments
- Probes
- 5mm Smartprobe with Z gradient. Broadband capable of 1H, 19F (simultaneous acquisition) and all frequencies from 31P-199Hg and 17O-109Ag
- Sundries
- Bruker 24 sample carosel
- Applications
- Automation software equipped with a 24 sample changer makes this ideal for characterization of small molecules and unusual nuclei. All 1D or 2D experiments based on 1H/X pairs such at DEPT, COSY, TOCSY, NOESY, ROESY, HSQC and HMBC are possible.