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(1)H NMR and isothermal titration calorimetry (ITC) experiments were employed to obtain reliable thermodynamic data for the formation of the 1:1 inclusion complexes of fullerenes C(60) and C(70) with the buckycatcher (C(60)H(28)). NMR measurements were done in toluene-d8 and chlorobenzene-d5 at 288, 298, and 308 K, while the ITC titrations were performed in toluene, chlorobenzene, o-dichlorobenzene, anisole, and 1,1,2,2-tetrachloroethane at temperatures from 278 to 323 K. The association constants, K(a), obtained with both techniques are in very good agreement. The thermodynamic data obtained by ITC indicate that generally the host-guest association is enthalpy-driven. Interestingly, the entropy contributions are, with rare exceptions, slightly stabilizing or close to zero. Neither ΔH nor ΔS is constant over the temperature range studied, and these thermodynamic functions exhibit classical enthalpy/entropy compensation. The ΔCp values calculated from the temperature dependence of the calorimetric ΔH values are negative for the association of both fullerenes with the buckycatcher in toluene. The negative ΔCp values are consistent with some desolvation of the host-cavity and the guest in the inclusion complexes, C(60)@C(60)H(28) and C(70)@C(60)H(28).

The specific recognition by proteins of G-quadruplex structures provides evidence of a functional role for in vivo G-quadruplex structures. As previously reported, the ribonucleoprotein, hnRNP Al, and it is proteolytic derivative, unwinding protein 1 (UP1), bind to and destabilize G-quadruplex structures formed by the human telomeric repeat d(TTAGGG)n. UP1 has been proposed to be involved in the recruitment of telomerase to telomeres for chain extension. In this study, a detailed thermodynamic characterization of the binding of UP1 to a human telomeric repeat sequence, the d[AGGG(TTAGGG)3] G-quadruplex, is presented and reveals key insights into the UP1-induced unfolding of the G-quadruplex structure. The UP1-G-quadruplex interactions are shown to be enthalpically driven, exhibiting large negative enthalpy changes for the formation of both the Na(+) and K(+) G-quadruplex-UP1 complexes (ΔH values of -43 and -19 kcal/mol, respectively). These data reveal three distinct enthalpic contributions from the interactions of UP1 with the Na(+) form of G-quadruplex DNA. The initial interaction is characterized by a binding affinity of 8.5 × 10(8) M(-1) (strand), 200 times stronger than the binding of UP1 to a single-stranded DNA with a comparable but non-quadruplex-forming sequence [4.1 × 10(6) M(-1) (strand)]. Circular dichroism spectroscopy reveals the Na(+) form of the G-quadruplex to be completely unfolded by UP1 at a binding ratio of 2:1 (UP1:G-quadruplex DNA). The data presented here demonstrate that the favorable energetics of the initial binding event are closely coupled with and drive the unfolding of the G-quadruplex structure.

Dissipative optomechanics has some advantages in cooling compared to the conventional dispersion dominated systems. Here, we study the optical response of a cantilever-like, silica, microsphere pendulum, evanescently coupled to a fiber taper. In a whispering gallery mode resonator the cavity mode and motion of the pendulum result in both dispersive and dissipative optomechanical interactions. This unique mechanism leads to an experimentally observable, asymmetric response function of the transduction spectrum which can be explained using coupled-mode theory. The optomechanical transduction, and its relationship to the external coupling gap, are investigated and we show that the experimental behavior is in good agreement with the theoretical predictions. A deep understanding of this mechanism is necessary to explore trapping and cooling in dissipative optomechanical systems., 5 pages

We have previously shown that c-MYC promoter sequences can form stable i-motifs in acidic solution (pH 4.5–5.5). In terms of drug targeting, the question is whether c-MYC promoter sequence i-motifs will exist in the nucleus at neutral pH. In this work, we have investigated the stability of a mutant c-MYC i-motif in solutions containing a molecular crowding agent. The crowded nuclear environment was modeled by the addition of up to 40% w/w polyethylene glycols having molecular weights up to 12,000 g/mol. CD and DSC were used to establish the presence and stability of c-MYC i-motifs in buffer solutions over the pH range 4 to 7. We have shown that the c-MYC i-motif can exist as a stable structure at pH values as high as 6.7 in crowded solutions. Generic dielectric constant effects, e.g., a shift in the pKa of cytosine by more than 2 units (e.g., 4.8 to 7.0), or the formation of non-specific PEG/DNA complexes appear to contribute insignificantly to i-motif stabilization. Molecular crowding, largely an excluded volume effect of added PEG, having a molecular weight in excess of 1,000 g/mol, appears to be responsible for stabilizing the more compact i-motif over the random coil at higher pH values.

Elastin-like Polypeptides (ELPs) are genetically engineered biopolymers that are derived from the endogenous protein Tropoelastin. ELPs are structurally disordered and soluble at low temperatures but transition to a β-spiral and aggregate at a Transition Temperature (TT). This aggregation is being explored as a novel drug delivery vector by thermally targeting systemically delivered ELP-drug conjugates. We are investigating the biophysical properties of ELP[V5G3A2-150] as a means of understanding and predicting the behavior in vivo. We have investigated the hydrodynamic, structural and thermodynamic properties of ELP[V5G3A2-150] through the use of CD, turbidity, DLS, DSC and SV. DLS and SV analyses suggest that ELP[V5G3A2-150] experiences small amounts of weak association below the TT that increase with temperature. CD analyses further indicate that below the TT ELP[V5G3A2-150] consists of both disordered (≈75%) and β-conformation (≈25%) and as the temperature and concentration is increased the % β-conformation increases. The temperature & concentration dependence of β-conformation suggests that the weak association can be attributed to heterogeneous β-sheets. SV revealed that above the TT ELP[V5G3A2-150] exhibits a temperature dependent critical concentration (CC). This CC is consistent with the TT and suggests that the TT may be described as a solubility constant.Assembly in serum raises the TT by ≈ 2.5°C. This is opposite to the expected effect of macromolecular crowding and suggests that certain serum proteins may be associating with ELP[V5G3A2-150]. Investigation of this effect through SV was greatly complicated by the presence of the Johnston-Ogston (J-O) effect. Further investigation suggested additional complexity in systems exhibiting the J-O effect than previously reported. Two of the additional complexities already determined are cross-term hydrodynamic non-ideality and high-concentration convection. Additional research into the effects of attaching CPPs to ELP[V5G3A2-150] will be presented. Work supported by NSF ARRA 0959211 grant.