RESUMO
Customization of deuterated biomolecules is vital for many advanced biological experiments including neutron scattering. However, because it is challenging to control the proportion and regiospecificity of deuterium incorporation in live systems, often only two or three synthetic lipids are mixed together to form simplistic model membranes. This limits the applicability and biological accuracy of the results generated with these synthetic membranes. Despite some limited prior examination of deuterating Escherichia coli lipids in vivo, this approach has not been widely implemented. Here, an extensive mass spectrometry-based profiling of E. coli phospholipid deuteration states with several different growth media was performed, and a computational method to describe deuterium distributions with a one-number summary is introduced. The deuteration states of 36 lipid species were quantitatively profiled in 15 different growth conditions, and tandem mass spectrometry was used to reveal deuterium localization. Regressions were employed to enable the prediction of lipid deuteration for untested conditions. Small-angle neutron scattering was performed on select deuterated lipid samples, which validated the deuteration states calculated from the mass spectral data. Based on these experiments, guidelines for the design of specifically deuterated phospholipids are described. This unlocks even greater capabilities from neutron-based techniques, enabling experiments that were formerly impossible.
Assuntos
Difração de Nêutrons , Fosfolipídeos , Deutério/química , Difração de Nêutrons/métodos , Escherichia coli/metabolismo , Espectrometria de Massas em TandemRESUMO
High-resolution imaging methods of the iridocorneal angle (ICA) will lead to enhanced understanding of aqueous humor outflow mechanisms and a characterization of the trabecular meshwork (TM) morphology at the cellular level will help to better understand glaucoma mechanics (e.g., cellular level biomechanics of the particulate glaucomas). This information will translate into immense clinical value, leading to more informed and customized treatment selection, and improved monitoring of procedural interventions that lower intraocular pressure (IOP). Given ICA anatomy, imaging modalities that yield intrinsic optical sectioning or 3D imaging capability will be useful to aid in the visualization of TM layers. This minireview examines advancements in imaging the ICA in high-resolution.