Q. Who can use the facility?
A. The facility is operated mainly for the benefit for researchers at the University of Connecticut. Access by researchers at other institutions or by commercial users is also available, with a different fee schedule.

Q. How can the facility help me?
A. The facility is mainly used to determine the three-dimensional structures of proteins and protein complexes in solution. Protein structures are frequently helpful for understanding the biological function of a protein. It is not uncommon for structural homology to exist where little or no sequence homology exists, so structure can sometimes reveal function. Structures are also useful for designing biochemical experiments, since they often reveal binding or active sites, epitopes, or exposed residues suitable for labeling. NMR can also be used to studied molecular dynamics or conformational change, and can be used to screen ligand libraries for molecules that bind specifically to a target protein.

Q. How do I gain access?
A. Contact the Director, Dr. Jeff Hoch (hoch (at) uchc.edu) or the facility manager Dr. Mark Maciejewski (markm (at) neuron.uchc.edu).

Q. Can my protein be studied by NMR?
A. High resolution NMR investigations are most feasible for proteins less than approximately 25 kDa in mass and soluble to around 0.5 mM. In certain cases it may be possible to investigate proteins or complexes of larger size or lower solubility. Membrane proteins are difficult to study using high-resolution methods, but new techniques are emerging that may be applicable. Because NMR studies usually require labeling the protein with the stable isotopes 15N, 13C, and sometimes 2H, an expression system suitable for growth in labeled media must be available. Proteins must be purified (typically >95% is required), folded, and at least marginally stable. Preliminary characterization by circular dichroism and thermal or solvent denaturation is recommended.

Q. How difficult/expensive is sample preparation for NMR?
A. Expression in E. coli is usually straightforward, and requires minimal media made with isotope-labeled sources of carbon and nitrogen (typically glucose and ammonium salts). In vitro folding may be required if the protein accumulates in inclusion bodies. Assuming a yield of 10 mg of purified and folded protein per liter of media, the cost of one NMR sample is around a few thousand $. A recipe for labeled minimal media can be found here.

Q. What if I don't have funds to pay for sample preparation or instrument time?
A. Contact the director, Jeff Hoch (hoch (at) uchc.edu), to inquire about instrument time grants or possible collaborations.