Biomolecular NMR Laboratory

Services Offered

  • User training
  • Consultation on proposal preparation
  • 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)

The Biomolecular NMR Laboratory maintains two high field NMR spectrometers 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:

   3D structure of effector domain of anti-microbial 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 Dr. Scott Hefty, Associate Professor of Molecular Biosciences at the University of Kansas, Lawrence, KS.  

 

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 Dr. Jeroen Roeloff, Assistant Professor in the Division of Biology at Kansas State University, Manhattan, KS.

 

 

This 1D-1H-NMR spectra of isolated Zn binding domain (RING) of cbl protein (above) was obtained in collaboration with Dr. 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.

 

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 HAS

 

 

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.

 

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.