RESUMEN
To watch biological macromolecules perform their functions inside the living cells is the dream of any biologists. In-cell nuclear magnetic resonance is a branch of biomolecular NMR spectroscopy that can be used to observe the structures, interactions and dynamics of these molecules in the living cells at atomic level. In principle, in-cell NMR can be applied to different cellular systems to achieve biologically relevant structural and functional information. In this review, we summarize the existing approaches in this field and discuss its applications in protein interactions, folding, stability and post-translational modifications. We hope this review will emphasize the effectiveness of in-cell NMR for studies of intricate biological processes and for structural analysis in cellular environments.
Asunto(s)
Sustancias Macromoleculares/química , Espectroscopía de Resonancia Magnética/métodos , Mapeo de Interacción de Proteínas/métodos , Proteínas/química , Sustancias Macromoleculares/metabolismo , Pliegue de Proteína , Procesamiento Proteico-Postraduccional , Estabilidad Proteica , Proteínas/genética , Relación Estructura-ActividadRESUMEN
The lipoprotein lipase (LPL) gene can be considered a functional candidate gene that regulates fatty acid composition. In this study, genetic associations between fatty acid composition and exonic single nucleotide polymorphisms (SNPs) in the LPL gene were examined using 612 Korean cattle. We investigated the relationship between unsaturated fatty acids and five novel SNPs (c.322G>A, c.329A>T, c.527T>G, c.988C>T and c.1591G>A), and confirmed that three polymorphic SNPs (c.322G>A, c.329A>T and c.1591G>A) were associated with fatty acid composition. Korean cattle with an AA genotype of c.322G>A, c.329A>T, and GA genotype of c.1591G>A had higher levels of monounsaturated fatty acids and carcass traits (P < 0.05). Our findings confirmed that three novel SNPs we identified in the LPL gene can affect fatty acid composition and carcass traits. Therefore, selection for AA and GA genotypes should be recommended to genetically improve beef quality and flavor.
Asunto(s)
Ácidos Grasos/genética , Estudios de Asociación Genética , Lipoproteína Lipasa/genética , Carne , Animales , Bovinos , Exones , Ácidos Grasos/química , Genotipo , Fenotipo , Polimorfismo de Nucleótido SimpleRESUMEN
The extreme toxicity of cyanide and its continued use in various industries have raised concerns over environmental contamination and, therefore, considerable attention has given to develop facile and sensitive methods of cyanide detection. In this study, we developed highly sensitive and straightforward methods of cyanide detection using eosin-labeled glutathionylcobalamin (E-GSCbl) containing fluorescent eosin-labeled glutathione (E-GSH) as the upper axial ligand to the cobalt. E-GSH fluorescence was strongly quenched in E-GSCbl. The E-GSH ligand of E-GSCbl was replaced specifically by cyanide, showing recovery of the E-GSH fluorescence. This profluorescent property of E-GSCbl enabled detection of cyanide in aqueous solutions, yielding a lower detection limit of 10â¯nM (0.26⯵gâ¯L-1). Moreover E-GSH exhibited strong luminescence under UV-light that was quenched in E-GSCbl, and this allowed naked-eye detection of cyanide at concentrations as low as 100â¯nM. This study demonstrates that profluorescent E-GSCbl is a highly sensitive cyanide chemosensor that can detect nanomolar concentrations of cyanide.