Effects of Ca 2+ on the Structure and Dynamics of PIP3 in Model Membranes Containing PC and PS.

Phosphatidylinositol phosphates (PIPs) are a family of seven different eukaryotic membrane lipids that have a large role in cell viability, despite their minor concentration in eukaryotic cellular membranes. PIPs tightly regulate cellular processes such as cellular growth, metabolism, immunity, and development through direct interactions with partner proteins. Understanding the biophysical properties of PIPs in the complex membrane environment is important to understand how PIPs selectively regulate a partner protein. Here we investigate the structure and dynamics of PIP3 in lipid bilayers that are simplified models of the natural membrane environment. We probe the effects of the anionic lipid phosphatidylserine (PS) and the divalent cation Ca 2+ . We use solution and solid-state 1 H, 31 P, and 13 C NMR all at natural abundance combined with MD simulations to characterize the structure and dynamics of PIPs. 1 H and 31 P 1D spectra show good resolution at high temperatures with isolated peaks in the headgroup, interfacial, and bilayer regions. Site specific assignment of these 1D reporters were made and used to measure the effects of Ca 2+ and PS. In particular, the resolved 31 P signals of the PIP3 headgroup allowed for extremely well localized information about PIP3 phosphate dynamics, which the MD simulations were able to help explain. Cross polarization kinetics provided additional site-specific dynamics measurements for the PIP3 headgroups.

3D-Printable centrifugal devices for biomolecular solid state NMR rotors.

The advent of magic angle spinning (MAS) rates exceeding 100 kHz has facilitated the acquisition of 1H-detected solid-state NMR spectra of biomolecules with high resolution. However, challenges can arise when preparing rotors for these experiments, due to the physical properties of biomolecular solid samples and the small dimensions of the rotors. In this study, we have designed 3D-printable centrifugal devices that facilitate efficient and consistent packing of crystalline protein slurries or viscous phospholipids into 0.7 mm rotors. We demonstrate the efficacy of these packing devices using 1H-detected solid state NMR at 105 kHz. In addition to devices for 0.7 mm rotors, we have also developed devices for other frequently employed rotor sizes and styles. We have made all our designs openly accessible, and we encourage their usage and ongoing development as a shared effort within the solid state NMR community.

Flexible Zn-MOF with Rare Underlying scu Topology for Effective Separation of C6 Alkane Isomers.

Adsorptive separation by porous solids provides an energy-efficient alternative for the purification of important chemical species compared to energy-intensive distillations. Particularly, the separation of linear hexane isomers from its branched counterparts is crucial to produce premium grade gasoline with high research octane number (RON). Herein, we report the synthesis of a new, flexible zinc-based metal-organic framework, [Zn5(µ3-OH)2(adtb)2(H2O)5·5 DMA] (Zn-adtb), constructed from a butterfly shaped carboxylate linker with underlying (4,8)-connected scu topology capable of separating the C6 isomers nHEX, 3MP, and 23DMB. The sorbate-sorbent interactions and separation mechanisms were investigated and analyzed through in situ FTIR, solid state NMR measurements and computational modeling. These studies reveal that Zn-adtb discriminates the nHEX/3MP isomer pair through a kinetic separation mechanism and the nHEX/23DMB isomer pair through a molecular sieving mechanism. Column breakthrough measurements further demonstrate the efficient separation of linear nHEX from the mono- and dibranched isomers.