We propose a thermodynamic model that includes the non-speci¢c binding of the V V phage regulatory proteins CI and Cro. By ¢tting the model to experimental in vivo data on activities of the two promoters P RM and P R versus concentration, we estimate the free energy upon non-speci¢c binding to be 3 34.1 þ 0.9 kcal/mol for CI and 3 34.2 þ 0.8 kcal/mol for Cro. For concentrations s 100 nM of CI or Cro, we ¢nd that s 50% of these proteins are non-specifically bound. In particular, in a lysogen (V V250 CI monomeric equivalents per cell) nearly 90% of CI is non-specifically bound. ß

We investigate the sensitivity of the right operator in bacteriophage lambda. In particular, the system is probed in the three different regulatory protein concentration-regimes: 1), lysogen (CI dominates); 2), during induction (CI and Cro at comparable concentrations); and 3), after induction (Cro dominates). Systematic perturbations of the protein-operator binding energies show in a lysogen that the activity (production rate) at promoter PRM is robust to variations, in contrast to PR, where the sensitivity is high. Both promoters, however, show large sensitivity in regimes 2 and 3. In all regimes we identify several suppressors, meaning that for a given large perturbation (+/-2 kcal/mol) of one binding energy, there exist compensating perturbation(s) that restore the wild-type activity.

Thermodynamic measurements of proteins indicate that the folding to the native state takes place either through stable intermediates or through a two-state process without intermediates. The rather short folding times of proteins indicate that folding is guided through some sequence of contact bindings. We discuss the possibility of reconciling a two-state folding event with a sequential folding process in a schematic model of protein folding. We propose a new dynamical transition temperature that is lower than the temperature at which proteins in equilibrium unfold. This is in qualitative agreement with observations of in vivo protein folding activity quantified by chaperone concentration in Escherichia coli. Finally, we discuss our framework in connection with the unfolding of proteins at low temperatures.

Thermodynamics related to hydrated water upon protein unfolding is studied over a broad temperature range (5-125 degrees C). The hydration effect arising from the apolar interior is modeled as an increased number of hydrogen bonds between water molecules compared with bulk water. The corresponding contribution from the polar interior is modeled as a two-step process. First, the polar interior breaks hydrogen bonds in bulk water upon unfolding. Second, due to strong bonds between the polar surface and the nearest water molecules, we assume quantization using a simplified two-state picture. The heat capacity change upon hydration is compared with model compound data evaluated previously for 20 different proteins. We obtain good correspondence with the data for both the apolar and the polar interior. We note that the effective coupling constants for both models have small variations among the proteins we have investigated.

We study a synthetic clay suspension of laponite at different particle and NaCl concentrations by measuring stationary shear viscosity and transient electrically induced birefringence (TEB). On one hand the viscosity data are consistent with the particles being spheres and the particles being associated with large amount bound water. On the other hand the viscosity data are also consistent with the particles being asymmetric, consistent with single laponite platelets associated with a very few monolayers of water. We analyze the TEB data by employing two different models of aggregate size (effective hydrodynamic radius) distribution: (1) bidisperse model and (2) log-normal distributed model. Both models fit, in the same manner, fairly well to the experimental TEB data and they indicate that the suspension consists of polydisperse particles. The models also appear to confirm that the aggregates increase in size vs increasing ionic strength. The smallest particles at low salt concentrations seem to be monomers and oligomers.