Determination of the dipole moments of RNAse SA wild type and a basic mutant.
In this study, we report the effects of acidic to basic residue point mutations (5K) on the dipole moment of RNAse SA at different pHs. Dipole moments were determined by measuring solution capacitance of the wild type (WT) and the 5K mutant with an impedance analyzer. The dipole moments were then (1) compared with theoretically calculated dipole moments, (2) analyzed to determine the effect of the point mutations, and (3) analyzed for their contribution to overall protein-protein interactions (PPI) in solution as quantitated by experimentally derived second virial coefficients. We determined that experimental and calculated dipoles were in reasonable agreement. Differences are likely due to local motions of residue side chains, which are not accounted for by the calculated dipole. We observed that the proteins' dipole moments increase as the pH is shifted further from their isoelectric points and that the wild-type dipole moments were greater than those of the 5K. This is likely due to an increase in the proportion of one charge (either negative or positive) relative to the other. A greater charge disparity corresponded to a larger dipole moment. Finally, the larger dipole moments of the WT resulted in greater attractive overall PPI for that protein as compared to the 5K.
Pharmaceutics, GPSC, Abbott Bioresearch Center, Worcester, Massachusetts 01605, USA.
Ravi
R
Chari
Shubhadra N
SN
Singh
Sandeep
S
Yadav
David N
DN
Brems
Devendra S
DS
Kalonia
D025802:Electric Capacitance; D006863:Hydrogen-Ion Concentration; D008239:Lysine / Q000737:chemistry; D056004:Molecular Dynamics Simulation; D050505:Mutant Proteins / Q000737:chemistry*; D009994:Osmolar Concentration; D017354:Point Mutation; D025941:Protein Interaction Mapping; D017434:Protein Structure, Tertiary; D012260:Ribonucleases / Q000737:chemistry* / Q000235:genetics; D012995:Solubility; D012996:Solutions / Q000737:chemistry; D055672:Static Electricity
D050505:Mutant Proteins; D012996:Solutions; D012260:Ribonucleases; C077002:ribonuclease Sa3; D008239:Lysine