Increasing the sensitivity of cryoprobe protein NMR experiments by using the sole low-conductivity arginine glutamate salt

Decrease in experimental sensitivity of cryoprobe experiments for salty samples, attributed to increased sample conductivity, has been a long-standing issue in protein NMR. Salt concentration can not be simply reduced as this often leads to protein aggregation. A simple and inexpensive solution to this problem is demonstrated here. We show that even for proteins prone to aggregation, the traditional solubilizing salt, 100mM NaCl, can be completely replaced by 50mM l-Arg and l-Glu. This replacement simultaneously reduces the sample conductivity and improves protein solubility. Up to a 6-fold overall increase in experimental sensitivity was achieved, in comparison with the traditional salty buffer. At constant protein concentration up to 2-fold increase in sensitivity was observed. The lengths of the proton pi/2 pulses were also significantly decreased, up to the level typical for non-salty samples in water.     

A simple method for improving protein solubility and long-term stability

Increasing a protein concentration in solution to the required level, without causing aggregation and precipitation is often a challenging but important task, especially in the field of structural biology; as little as 20% of nonmembrane proteins have been found to be suitable candidates for structural studies predominantly due to poor protein solubility. We demonstrate here that simultaneous addition of charged amino acids L-Arg and L-Glu at 50 mM to the buffer can dramatically increase the maximum achievable concentration of soluble protein (up to 8.7 times). These amino acids are effective in preventing protein aggregation and precipitation, and they dramatically increase the long-term stability of the sample; additionally, they protect protein samples from proteolytic degradation. Specific protein-protein and protein-RNA interactions are not adversely affected by the presence of these amino acids. These additives are particularly suitable for situations where high protein concentration and long-term stability are required, including solution-state studies of isotopically labeled proteins by NMR.     

Association of Molybdenum Ionic Species with Alumina Surface

The adsorption isotherms at 25, 45, and 65 degrees C of molybdenum solutions of concentration ranges between 10(-3) and 3x10(-2) M(Mo) (pH 4-5) on different alumina samples are investigated. The analysis is conducted using a modified Frumkin isotherm which takes a more realistic account of the lateral interaction between adsorbed species and considers that the adsorption takes place on the most basic OH groups on the surface of alumina. The results are discussed in view of the difference in solutions speciation, and the changes in the pH of the remaining supernatant solutions. The solution temperature, PZC of the used aluminas, the configuration of the basic OH groups on their surface, and the pore structure have been shown to intervene effectively. Copyright 2000 Academic Press.