A large agricultural company collaborated with the David H. Murdock Research Institute (DHMRI) to determine the in-solution three dimensional protein structure of a 275-residue, DNA-binding domain of a double labeled (13C, 15N) protein.
To determine the protein structure, both 700 MHz and 950 MHz NMR spectrometers were used to perform a variety of three dimensional experiments that determined chemical shift assignments of atoms within the protein. To assign chemical shift resonance, DHMRI scientists used NMRPipe and NMRView software. The DHMRI’s 950 MHz NMR spectrometer was also used for 3D NOESY experiments. Structure was determined using Cyana software and NOESY values for constraints for energy minimization. AMBER software was used to refine structure in water environment.
The project highlights the expertise of DHMRI scientists to apply an array of NMR services to determine a protein sub-domain and provided collaborators with detailed insight into DNA binding.