Electrostatic Contributions to the Thermodynamics of Ribonuclease P Protein Folding

Electrostatic interactions are of fundamental importance in determining the structure and stability of macromolecules. For example, charge-charge interactions modulate the folding and binding of proteins and influence protein solubility. Electrostatic interactions are highly variable and can be both favorable and unfavorable. The ability to quantify these interactions is challenging but vital to understanding the detailed balance and major roles that they have in different proteins and biological processes. Measuring pKa values of ionizable groups provides a sensitive method for experimentally probing the electrostatic properties of a protein.

pKa values report the free energy of site-specific proton binding and provide a direct means of studying protein folding and pH-dependent stability. Using a combination of NMR, circular dichroism, and fluorescence spectroscopy along with singular value decomposition, we investigated the contributions of electrostatic interactions to the thermodynamic stability and folding of the protein subunit of Bacillus subtilis ribonuclease P, P protein. Taken together, the results suggest that unfavorable electrostatics alone do not account for the fact that P protein is intrinsically unfolded in the absence of ligand because the pKa differences observed between the folded and unfolded state are small. Presumably, multiple factors encoded in the P protein sequence account for its IUP property, which may play an important role in its function.

Common Bound: The Small Groups of Methodism

The system of small groups John Wesley established to promote a proper life of discipleship in early Methodist converts was, in many respects, the strength of the Methodist movement. Those who responded to Wesley’s initial invitation to “flee the wrath to come” were organized into large gatherings called “societies,” which were then subdivided into smaller bands, class meetings, select societies, and penitent bands. The smaller groups gave Wesley the opportunity, through a system of appointed leaders, to keep track of the spiritual progress of every member in his movement, which grew to tens of thousands by the time of his death in 1791. As Methodism shifted from renewal movement to institutional church in the nineteenth century, however, growth slowed, and participation in such groups declined rapidly. By the early twentieth century, classes and bands were virtually extinct in every sector of Methodism save the African-American tradition. In recent years, scholars in various sectors of the Wesleyan tradition, particularly David Lowes Watson and Kevin Watson, have called for a recovery of these small groups for purposes of renewal in the church. There is no consensus, however, concerning what exactly contributed to the vitality of these groups during Wesley’s ministry.

Over the last century, sociological studies of group dynamics have revealed three common traits that are crucial to highly functioning groups: interdependence created by the existence of a common goal, interaction among group members that is “promotive” or cooperative in nature, and high levels of feedback associated with personal responsibility and individual accountability. All three of these were prevalent in the early Methodist groups. Interdependence existed around a shared goal, which for Wesley and the Methodists was holiness. That interdependence was cooperative in nature; individuals experienced the empowering grace of God as they each pursued the goal in the company of fellow pilgrims. Finally, the groups existed for purposes of feedback and accountability as individuals took responsibility both for themselves and others as they progressed together toward the goal of holy living. Wesley seemed to instinctively understand the essential nature of each of these characteristics in maintaining the vitality of the movement when he spoke of the importance of preserving the “doctrine, spirit and discipline” of early Methodism. Analysis of some of the present-day attempts to restore Wesley’s groups reveals frequent neglect to one or more of these three components. Perhaps most critical to recovering the vitality of the early Methodist groups will be reclaiming the goal of sanctification and coming to a consensus on what its pursuit means in the present day.