Biomedical subjects

S Garofoli

Publications and source records attributed to S Garofoli.

3 recordsLinked to original sources

PubMed2004

Thermophilicity of wild type and mutant cold shock proteins by molecular dynamics simulation.

In the attempt to clarify possible mechanisms underlying thermal stability of proteins, we study through molecular dynamics thermophile Bc-Csp, mesophile Bs-CspB, and selected mutants. These proteins have been extensively characterized experimentally; researchers showed that differential thermostability among the wild type proteins is fundamentally linked to one or two mutated amino acids, and that the nature of the effect is electrostatic. They also inferred an atomistic mechanism related to removal of unfavorable interactions, rather than to the formation of salt bridges. Molecular dynamics allows us to confirm and support both hypotheses. Several other collective parameters have also been monitored in relation to thermophilicity, such as global and local rigidity, permanence and number of hydrogen bonds, or of salt links. None of these clearly correlates with the thermal stability of the presently studied proteins.

Bacillus↗

PubMed2003

Modeling permeation energetics in the KcsA potassium channel.

The thermodynamics of cation permeation through the KcsA K(+) channel selectivity filter is studied from the perspective of a physically transparent semimicroscopic model using Monte Carlo free energy integration. The computational approach chosen permits dissection of the separate contributions to ionic stabilization arising from different parts of the channel (selectivity filter carbonyls, single-file water, cavity water, reaction field of bulk water, inner helices, ionizable residues). All features play important roles; their relative significance varies with the ion's position in the filter. The cavity appears to act as an electrostatic buffer, shielding filter ions from structural changes in the inner pore. The model exhibits K(+) vs. Na(+) selectivity, and roughly isoenergetic profiles for K(+) and Rb(+), and discriminates against Cs(+), all in agreement with experimental data. It also indicates that Ba(2+) and Na(+) compete effectively with permeant ions at a site near the boundary between the filter and the cavity, in the vicinity of the barium blocker site.

Bacterial Proteins↗

PubMed1999

Ionic interactions in multiply occupied channels.

A significant number of physiologically important ion channels function via multi-ion mechanisms where repulsion between ions at slightly separated locations is believed to be critical for permeation. We apply the semi-microscopic Monte Carlo approach and analyse how multiple occupancy affects permeation energetics and ion-water-peptide correlations. We consider double occupancy in idealized models of two systems: gramicidin A and the KcsA K+ channel. We focus on the excess repulsion energy due to ion-water and ion-peptide correlations (repulsion energy adjusted for direct ion-ion interaction). Gramicidin, where multiple occupancy is marginally important functionally, is ideal for correlating structure and ion interactions. Pair occupancy is stabilized by interaction with bulk solvent, destabilized by interaction with both the channel water and, as binding sites are far apart, the peptide backbone. In the KcsA K+ channel, double occupancy is promoted by the uneven spacing and the large ion-water separations in the selectivity filter. The carbonyls forming the binding cavities are equally important for pair stabilization. Due to the binding pocket's design, net ionic repulsion is approximately 25-30% of what it would be in a gramicidin-like structure with the same interionic spacing.

Anti-Bacterial Agents↗