Determination of fatty acid uptake and desaturase activity in mammalian cells by NMR-based stable isotope tracing.
Anal Chim Acta
; 1303: 342511, 2024 May 15.
Article
in En
| MEDLINE
| ID: mdl-38609261
ABSTRACT
BACKGROUND:
Mammalian cells both import exogenous fatty acids and synthesize them de novo. Palmitate, the end product of fatty acid synthase (FASN) is a substrate for stearoyl-CoA desaturases (Δ-9 desaturases) that introduce a single double bond into fatty acyl-CoA substrates such as palmitoyl-CoA and stearoyl-CoA. This process is particularly upregulated in lipogenic tissues and cancer cells. Tracer methodology is needed to determine uptake versus de novo synthesis of lipids and subsequent chain elongation and desaturation. Here we describe an NMR method to determine the uptake of 13C-palmitate from the medium into HCT116 human colorectal cancer cells, and the subsequent desaturation and incorporation into complex lipids.RESULTS:
Exogenous 13C16-palmitate was absorbed from the medium by HCT116 cells and incorporated primarily into complex glycerol lipids. Desaturase activity was determined from the quantification of double bonds in acyl chains, which was greatly reduced by ablation of the major desaturase SCD1.SIGNIFICANCE:
The NMR approach requires minimal sample preparation, is non-destructive, and provides direct information about the level of saturation and incorporation of fatty acids into complex lipids.Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Magnetic Resonance Imaging
/
Bisphenol A-Glycidyl Methacrylate
/
Fatty Acids
Limits:
Animals
/
Humans
Language:
En
Journal:
Anal Chim Acta
Year:
2024
Type:
Article
Affiliation country:
United States