Membrane remodelling plays an important role in cellular tasks such as endocytosis, vesiculation and protein sorting, and in the biogenesis of organelles such as the endoplasmic reticulum or the Golgi apparatus. It is well established that the remodelling process is aided by specialized proteins that can sense as well as create membrane curvature, and trigger tubulation when added to synthetic liposomes. Because the energy needed for such large-scale changes in membrane geometry significantly exceeds the binding energy between individual proteins and between protein and membrane, cooperative action is essential. It has recently been suggested that curvature-mediated attractive interactions could aid cooperation and complement the effects of specific binding events on membrane remodelling. But it is difficult to experimentally isolate curvature-mediated interactions from direct attractions between proteins. Moreover, approximate theories predict repulsion between isotropically curving proteins. Here we use coarse-grained membrane simulations to show that curvature-inducing model proteins adsorbed on lipid bilayer membranes can experience attractive interactions that arise purely as a result of membrane curvature. We find that once a minimal local bending is realized, the effect robustly drives protein cluster formation and subsequent transformation into vesicles with radii that correlate with the local curvature imprint. Owing to its universal nature, curvature-mediated attraction can operate even between proteins lacking any specific interactions, such as newly synthesized and still immature membrane proteins in the endoplasmic reticulum.
Vesicles formed in water by synthetic macro-amphiphiles have attracted much attention as nanocontainers having properties that extend the physical and chemical limits of liposomes. We sought to develop ABA block copolymeric amphiphiles that self-assemble into unilamellar vesicles that can be further oxidatively destabilized. We selected poly(ethylene glycol) (PEG) as the hydrophilic A blocks, owing to its resistance to protein adsorption and low toxicity. As hydrophobic B blocks, we selected poly(propylene sulphide) (PPS), owing to its extreme hydrophobicity, its low glass-transition temperature, and most importantly its oxidative conversion from a hydrophobe to a hydrophile, poly(propylene sulphoxide) and ultimately poly(propylene sulphone). This is the first example of the use of oxidative conversions to destabilize such carriers. This new class of oxidation-responsive polymeric vesicles may find applications as nanocontainers in drug delivery, biosensing and biodetection.
In this paper, we present the application of four different in situ analytical techniques to monitor the solvent-mediated polymorphic transformation of L-glutamic acid. Focused beam reflectance measurement (FBRM) and particle vision and measurement (PVM) have been used to track the chord length and morphology of the crystals over the course of the transformation. The polymorphic forms present have been monitored using Raman spectroscopy, while attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy has been used to measure the liquid-phase concentration profile. The combination of the different in situ data was used to identify the fundamental phenomena of nucleation and growth that govern the process. Moreover, the measurement data were combined with a mathematical model based on population balance equations and the fundamental equations describing the kinetics of nucleation and growth of both polymorphs. This combination allowed for the estimation of the characteristic nucleation and growth rates of the two polymorphic forms, while the dissolution process of the metastable polymorph was estimated using a Sherwood correlation. Finally, the experimental results obtained with different initial conditions and their simulation allowed for the validation of the population balance model and for a deeper understanding of the transformation process.
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