Video Experimental Relacionado
Updated: May 18, 2026
10:25
Monitoring Protein-Ligand Interactions in Human Cells by Real-Time Quantitative In-Cell NMR using a High Cell Density Bioreactor
Published on: March 9, 2021
On-Cell MAS NMR: pistas fisiológicas de las células vivas
Giorgia Zandomeneghi1, Karin Ilg, Markus Aebi
1Physical Chemistry, ETH Zurich, Wolfgang-Pauli Str. 10, 8093 Zurich, Switzerland.
Journal of the American Chemical Society
|October 4, 2012
Resumen
Los investigadores estudiaron la Salmonella enterica serovar Typhimurium viva mediante el uso de la espectroscopia de resonancia magnética nuclear (RMN). Observaron cambios en el antígeno O en la membrana externa bacteriana, incluido su estado de O-acetilación, durante la maduración.
Área de la Ciencia:
- Microbiología y Bioquímica.
- Química biofísica y bioquímica.
- Biología Estructural Biología estructural.
Sus antecedentes:
- La información estructural sobre las biomoléculas se deriva típicamente de muestras purificadas in vitro.
- La espectroscopia de Resonancia Magnética Nuclear (RMN) ofrece el potencial para analizar biomoléculas dentro de células o organismos intactos.
- Salmonella enterica serovar Typhimurium es una causa importante de la gastroenteritis en el hombre.
Objetivo del estudio:
- Para investigar los procesos de maduración en Salmonella enterica serovar Typhimurium vivo.
- Para controlar la composición del antígeno O en la membrana externa bacteriana durante la maduración.
- Evaluar el papel del estado de O-acetilación en la composición del antígeno O y su impacto potencial en el desarrollo de vacunas.
Principales métodos:
- Se empleó la espectroscopia de RMN de giro de ángulo mágico (MAS) de alta resolución.
- El estudio se centró en el análisis fisiológico de células bacterianas vivas.
- Se realizó la caracterización de la composición del antígeno O y el estado de O-acetilación.
Principales resultados:
- El estudio siguió con éxito la composición del antígeno O en Salmonella viva.
- Se detectó y caracterizó una evolución de la composición del antígeno O.
- Se identificaron específicamente cambios en el estado de O-acetilación del antígeno O.
Conclusiones:
- La espectroscopia de RMN es un método viable para estudiar los procesos de maduración en bacterias enteras.
- La composición del antígeno O, particularmente su estado de O-acetilación, evoluciona durante la maduración de Salmonella.
- Comprender estos cambios es crucial para las posibles estrategias de desarrollo de vacunas contra las infecciones por Salmonella.
Videos de Conceptos Relacionados
01:25Applications Of NMR In Biology
Nuclear magnetic resonance (NMR) spectroscopy is a very valuable analytical technique for researchers. It has been used for more than 50 years as an analytical tool. F. Bloch and E. Purcell formulated NMR in 1946 and won the 1952 Nobel Prize in Physics for their work. Biological macromolecules such as proteins, nucleic acids, lipids, and organic molecules including pharmaceutical compounds, can be studied using this versatile tool that exploits the magnetic properties of certain nuclei.
The...
The...
01:07Nuclear Magnetic Resonance (NMR): Overview
Nuclear magnetic resonance (NMR) is a phenomenon exhibited by certain nuclei that can absorb characteristic radio frequency radiation under certain conditions. NMR has been extensively applied in molecular spectroscopy and medical diagnostic imaging. In both these applications, the molecule or subject under study is placed in a magnetic field and irradiated with radio frequency energy.
NMR spectroscopy generates a spectrum where the characteristic absorption frequencies of the sample are...
NMR spectroscopy generates a spectrum where the characteristic absorption frequencies of the sample are...
01:16Calmodulin-dependent Signaling
Calmodulin (CaM) is a calcium-binding protein in eukaryotes that controls various calcium-regulated cellular processes. It has four calcium-binding sites that bind calcium to form the calcium-calmodulin ( Ca2+-CaM) complex. GPCR stimulation increases the calcium levels in the cells that bind to CaM and induces a conformational change.
The Ca2+-CaM complex does not have enzymatic activity by itself. Instead, the complex binds downstream target proteins, including membrane proteins or enzymes,...
The Ca2+-CaM complex does not have enzymatic activity by itself. Instead, the complex binds downstream target proteins, including membrane proteins or enzymes,...
01:19Protein Dynamics in Living Cells
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
01:162D NMR: Overview of Homonuclear Correlation Techniques
Homonuclear correlation spectroscopy (COSY) is a powerful technique used in Nuclear Magnetic Resonance (NMR) spectroscopy to study the correlations between nuclei of the same type within a molecule. It provides information about scalar couplings between adjacent nuclei, which helps determine connectivity and structural information. There are several COSY variants, each with its unique strengths and experimental parameters.
COSY90 is the standard two-dimensional (2D) COSY experiment that...
COSY90 is the standard two-dimensional (2D) COSY experiment that...
01:17Chemical Shift: Internal References and Solvent Effects
In an NMR sample, precise measurement of the absolute absorption frequencies of nuclei is difficult. A standard internal reference compound is added, and the frequency difference between the reference signal and sample signals is measured.
The internal reference compound generally used in NMR spectroscopy is tetramethylsilane (TMS). TMS is preferred because it is chemically inert, soluble in NMR solvents, and easily removable. Also, the highly shielded methyl protons in TMS yield an intense...
The internal reference compound generally used in NMR spectroscopy is tetramethylsilane (TMS). TMS is preferred because it is chemically inert, soluble in NMR solvents, and easily removable. Also, the highly shielded methyl protons in TMS yield an intense...