Cooperativity during Melting and Molecular Exchange in Micelles with Crystalline Cores

Molecular exchange processes are important equilibration and transport mechanisms in both synthetic and biological self-assembled systems such as micelles, vesicles, and membranes. Still, these processes are not entirely understood, in particular the effect of crystallinity and the interplay between cooperative melting processes and chain exchange. Here we focus on a set of simple polymer micelles formed by binary mixtures of poly(ethylene oxide)-mono-n-alkyl-ethers (Cn−PEO5) which allows the melting point to be tuned over a wide range. We show that the melting transition is cooperative in the confined 4–5 nm micellar core, whereas the exchange process is widely decoupled and unimeric in nature. As confirmed by differential scanning calorimetry, the total activation energy for ejecting a molecule out of the micellar core below the melting point is the sum of the enthalpy of fusion and the corresponding activation energy in the melt state. This suggests that a “local, single-chain melting process” preludes the molecular diffusion out of the micelle during chain exchange

Spherical Micelles with Nonspherical Cores: Effect of Chain Packing on the Micellar Shape

Self-assembly of amphiphilic polymers into micelles is an archetypical example of a "self-confined"system due to the formation of micellar cores with dimensions of a few nanometers. In this work, we investigate the chain packing and resulting shape of Cn-PEOx micelles with semicrystalline cores using small/wide-angle X-ray scattering (SAXS/WAXS), contrast-variation small-angle neutron scattering (SANS), and nuclear magnetic resonance spectroscopy (NMR). Interestingly, the n-alkyl chains adopt a rotator-like conformation and pack into prolate ellipses (axial ratio ϵ ≈ 0.5) in the "crystalline"region and abruptly arrange into a more spheroidal shape (ϵ ≈ 0.7) above the melting point. We attribute the distorted spherical shape above the melting point to thermal fluctuations and intrinsic rigidity of the n-alkyl blocks. We also find evidence for a thin dehydrated PEO layer (≤1 nm) close to the micellar core. The results provide substantial insight into the interplay between crystallinity and molecular packing in confinement and the resulting overall micellar shape

Biophysical studies on interactions and assembly of full-size E3 ubiquitin ligase : suppressor of cytokine signaling 2 (SOCS2)-elongin BC-cullin 5-ring box protein 2 (RBX2)

The multisubunit cullin RING E3 ubiquitin ligases (CRLs) target post-translationally modified substrates for ubiquitination and proteasomal degradation. The suppressors of cytokine signaling (SOCS) proteins play important roles in inflammatory processes, diabetes, and cancer and therefore represent attractive targets for therapeutic intervention. The SOCS proteins, among their other functions, serve as substrate receptors of CRL5 complexes. A member of the CRL family, SOCS2-EloBC-Cul5-Rbx2 (CRL5(SOCS2)), binds phosphorylated growth hormone receptor as its main substrate. Here, we demonstrate that the components of CRL5(SOCS2) can be specifically pulled from K562 human cell lysates using beads decorated with phosphorylated growth hormone receptor peptides. Subsequently, SOCS2-EloBC and full-length Cul5-Rbx2, recombinantly expressed in Escherichia coli and in Sf21 insect cells, respectively, were used to reconstitute neddylated and unneddylated CRL5(SOCS2) complexes in vitro. Finally, diverse biophysical methods were employed to study the assembly and interactions within the complexes. Unlike other E3 ligases, CRL5(SOCS2) was found to exist in a monomeric state as confirmed by size exclusion chromatography with inline multiangle static light scattering and native MS. Affinities of the protein-protein interactions within the multisubunit complex were measured by isothermal titration calorimetry. A structural model for full-size neddylated and unneddylated CRL5(SOCS2) complexes is supported by traveling wave ion mobility mass spectrometry data

Biophysical studies on interactions and assembly of full-size E3 ubiquitin ligase:suppressor of cytokine signaling 2 (SOCS2):ElonginBC:Cullin5:RING-box protein 2 (Rbx2)

The multisubunit cullin RING E3 ubiquitin ligases (CRLs) target post-translationally modified substrates for ubiquitination and proteasomal degradation. The suppressors of cytokine signaling (SOCS) proteins play important roles in inflammatory processes, diabetes, and cancer and therefore represent attractive targets for therapeutic intervention. The SOCS proteins, among their other functions, serve as substrate receptors of CRL5 complexes. A member of the CRL family, SOCS2-EloBC-Cul5-Rbx2 (CRL5(SOCS2)), binds phosphorylated growth hormone receptor as its main substrate. Here, we demonstrate that the components of CRL5(SOCS2) can be specifically pulled from K562 human cell lysates using beads decorated with phosphorylated growth hormone receptor peptides. Subsequently, SOCS2-EloBC and full-length Cul5-Rbx2, recombinantly expressed in Escherichia coli and in Sf21 insect cells, respectively, were used to reconstitute neddylated and unneddylated CRL5(SOCS2) complexes in vitro. Finally, diverse biophysical methods were employed to study the assembly and interactions within the complexes. Unlike other E3 ligases, CRL5(SOCS2) was found to exist in a monomeric state as confirmed by size exclusion chromatography with inline multiangle static light scattering and native MS. Affinities of the protein-protein interactions within the multisubunit complex were measured by isothermal titration calorimetry. A structural model for full-size neddylated and unneddylated CRL5(SOCS2) complexes is supported by traveling wave ion mobility mass spectrometry data

How Can Data Drive Policy and Practice in Child Welfare? Making the Link in Canada

Formal university–child welfare partnerships offer a unique opportunity to begin to fill the gaps in the child welfare knowledge base and link child welfare services to the realities of practice. With resources from a knowledge mobilization grant, a formal partnership was developed between the University of Toronto, clinicians, policy analysts, and researchers from child welfare agencies across Ontario. The key objectives of the grant included: (1) enhancing the capacity of service providers to access and analyze child welfare data to inform service and policy decisions; (2) integrating clinical expertise in service and policy decisions; and (3) developing a joint research agenda addressing high-priority knowledge gaps. This partnership was an opportunity to advance the evidence base with respect to service provision in Ontario and to create a culture of knowledge and evidence that would eventually support more complex research initiatives. Administrative data was analyzed for this partnership through the Ontario Child Abuse and Neglect Data System (OCANDS)—the first child welfare data system in Ontario to track child welfare-involved children and their families. Child welfare agencies identified recurrence as an important priority and agency-driven analyses were subsequently conducted on OCANDS generated recurrence Service Performance Indicators (SPI’s). Using an urgent versus chronic investigative taxonomy for analyses, findings revealed that the majority of cases did not recur within 12 months and cases identified as chronic needs are more likely to return to the attention of child welfare authorities. One of the key outcomes of the partnership — helping agencies to understand their administrative data is described, as are considerations for next steps for future partnerships and research