Rapid two-dimensional ALSOFAST-HSQC experiment for metabolomics and fluxomics studies: application to a C-13-enriched cancer cell model treated with gold nanoparticles

Autores de IIS La Fe

• 

  Antonio Pineda Lucena

  Autor

Participantes ajenos a IIS La Fe

• Schatzlein, MP
• Becker, J
• Schulze-Sunninghausen, D
• Herance, JR
• Luy, B

Grupos

• Unidad de Descubrimiento de Fármacos (UDF)

  Investigador Principal

  Leonor Puchades Carrasco

Abstract

Isotope labeling enables the use of C-13-based metabolomics techniques with strongly improved resolution for a better identification of relevant metabolites and tracing of metabolic fluxes in cell and animal models, as required in fluxomics studies. However, even at high NMR-active isotope abundance, the acquisition of one-dimensional C-13 and classical two-dimensional H-1,C-13-HSQC experiments remains time consuming. With the aim to provide a shorter, more efficient alternative, herein we explored the ALSOFAST-HSQC experiment with its rapid acquisition scheme for the analysis of C-13-labeled metabolites in complex biological mixtures. As an initial step, the parameters of the pulse sequence were optimized to take into account the specific characteristics of the complex samples. We then applied the fast two-dimensional experiment to study the effect of different kinds of antioxidant gold nanoparticles on a HeLa cancer cell model grown on C-13 glucose-enriched medium. As a result, H-1,C-13-2D correlations could be obtained in a couple of seconds to few minutes, allowing a simple and reliable identification of various C-13-enriched metabolites and the determination of specific variations between the different sample groups. Thus, it was possible to monitor glucose metabolism in the cell model and study the antioxidant effect of the coated gold nanoparticles in detail. Finally, with an experiment time of only half an hour, highly resolved H-1,C-13-HSQC spectra using the ALSOFAST-HSQC pulse sequence were acquired, revealing the isotope-position-patterns of the corresponding C-13-nuclei from carbon multiplets.

Keywords

• Fast NMR; HSQC; Metabolomics; Fluxomics; Gold nanoparticles