Biomolecular NMR Laboratory

Laboratory staff provide advice, consultation, training, assistance and service to investigators. The staff is also responsible for maintenance and upgrades of the high field NMR instruments, implementation of new NMR pulse sequences, and assisting local and remote users with technical problems. The laboratory is equipped with a Bruker Avance III 600 MHz with a variety of probes. It is located in close proximity to other core service laboratories in the KU Structural Biology Center.

Services Offered

NMR data collection (1D to 3D/4D)
NMR data processing
NMR data analysis
- backbone sequence specific resonance assignment
- solve high resolution solution NMR structures
Biomolecular interaction studies by NMR
- protein-protein, protein-nucleic acid, protein-small molecule
Protein dynamics studies
- pico- to nano sec. time scale motion (T1, T2 and het-NOE)
- model-free analysis
- micro- to milli sec. time scale motions by CPMG
  - relaxation dispersion data analysis
Fragment screening
- saturation transfer difference (STD) NMR
- surface plasmon resonance (SPR)
User training
Consultation on proposal preparation

The Biomolecular NMR Laboratory maintains a high field NMR spectrometer in support of structural and dynamics studies of biomolecules. Its capabilities include determining high resolution structures, biological macromolecules, elucidation and structural mapping of protein-protein, protein-nucleic acid, protein-peptide, protein-drug interactions, and studies of the dynamics of proteins and their complexes in solution. Those projects have resulted in the following:

Protein ChxR

This graphic shows a three-dimensional structure of effector domain of the anti-microbial protein ChxR. The structure was solved by solution NMR technique performed in collaboration with Scott Hefty, Professor of Molecular Biosciences at the University of Kansas, Lawrence, KS.

3D structure of protein from proteasome

A three dimensional structure of 10 kDa protein from proteasome regulatory particle was solved by solution NMR technique. This work was conducted in collaboration with Jeroen Roeloff, Associate Professor in the Division of Biology at Kansas State University, Manhattan, KS.

1D-1H-NMR spectra of isolated Zn binding domain (RING) of cbl protein

This 1D-1H-NMR spectra of isolated Zn binding domain (RING) of cbl protein (above) was obtained in collaboration with Raymond Perez, Professor, Clinical Research Center at the University of Kansas Medical Center. Upon addition of reducing agent the protein undergoes conformational change from unstructured to structured state as evidenced by the appearance of new peaks in the amide and alpha proton region (highlighted in oval shape) that are characteristic of formation of beta strand/sheet.

STD NMR

A. 600 MHz (BNMR core)1H-NMR spectrum with WATERGATE water suppression of 500 uM salicylic acid and 500 uM sucrose in 50 mM sodium phosphate, pH 7 (99.9% D2O), 0.1 mM sodium azide and 0.1 mM 4,4-dimethyl-4-silapentane-1-sulfonic acid (DSS).

B. Same as A, except in the presence of 10 uM human serum albumin (HSA)

C. STD spectrum showing that only salicylic acid is bound to human serum albumin (HSA)

2D-1H-13C(natural abundance)-HSQC-TOCSY spectrum

A 2D-1H-13C(natural abundance)-HSQC-TOCSY spectrum of small peptide was acquired using an 800 MHz NMR. By just using this spectrum alone researchers can easily assign all the proton and carbon atoms and their connectivity present in the chemical structure.

2D-1H-13C (natural abundance)-HSQC spectrum of small peptide

A 2D-1H-13C(natural abundance)-HSQC spectrum of small peptide is shown to the right. This data was acquired using an 800 MHz NMR.