» Articles » PMID: 26150938

Simultaneous Extraction of Proteins and Metabolites from Cells in Culture

Overview
Journal MethodsX
Specialty Pathology
Date 2015 Jul 8
PMID 26150938
Citations 64
Authors
Affiliations
Soon will be listed here.
Abstract

Proper sample preparation is an integral part of all omics approaches, and can drastically impact the results of a wide number of analyses. As metabolomics and proteomics research approaches often yield complementary information, it is desirable to have a sample preparation procedure which can yield information for both types of analyses from the same cell population. This protocol explains a method for the separation and isolation of metabolites and proteins from the same biological sample, in order for downstream use in metabolomics and proteomics analyses simultaneously. In this way, two different levels of biological regulation can be studied in a single sample, minimizing the variance that would result from multiple experiments. This protocol can be used with both adherent and suspension cell cultures, and the extraction of metabolites from cellular medium is also detailed, so that cellular uptake and secretion of metabolites can be quantified. Advantages of this technique includes:1.Inexpensive and quick to perform; this method does not require any kits.2.Can be used on any cells in culture, including cell lines and primary cells extracted from living organisms.3.A wide variety of different analysis techniques can be used, adding additional value to metabolomics data analyzed from a sample; this is of high value in experimental systems biology.

Citing Articles

A Dive Into Yeast's Sugar Diet-Comparing the Metabolic Response of Glucose, Fructose, Sucrose, and Maltose Under Dynamic Feast/Famine Conditions.

Verhagen K, Pardijs I, van Klaveren H, Wahl S Biotechnol Bioeng. 2025; 122(4):1035-1050.

PMID: 39865609 PMC: 11895419. DOI: 10.1002/bit.28935.


The potential therapeutic role of itaconate and mesaconate on the detrimental effects of LPS-induced neuroinflammation in the brain.

Ohm M, Hosseini S, Lonnemann N, He W, More T, Goldmann O J Neuroinflammation. 2024; 21(1):207.

PMID: 39164713 PMC: 11337794. DOI: 10.1186/s12974-024-03188-3.


Nitrate promotes the growth and the production of short-chain fatty acids and tryptophan from commensal anaerobe in the lactate-deficient environment by facilitating the catabolism of glutamate and aspartate.

Hung J, Zhang S, Huang S Appl Environ Microbiol. 2024; 90(8):e0114824.

PMID: 39082806 PMC: 11337843. DOI: 10.1128/aem.01148-24.


Optimizing MS-Based Multi-Omics: Comparative Analysis of Protein, Metabolite, and Lipid Extraction Techniques.

Mok J, Joo M, Cho S, Duong V, Song H, Park J Metabolites. 2024; 14(1).

PMID: 38248837 PMC: 10820684. DOI: 10.3390/metabo14010034.


Electronic cigarette vapor disrupts key metabolic pathways in human lung epithelial cells.

Assiri M, Al Jumayi S, Alsuhaymi S, Emwas A, Jaremko M, Alsaleh N Saudi Pharm J. 2023; 32(1):101897.

PMID: 38090735 PMC: 10714235. DOI: 10.1016/j.jsps.2023.101897.


References
1.
Ewald J, Heux S, Zamboni N . High-throughput quantitative metabolomics: workflow for cultivation, quenching, and analysis of yeast in a multiwell format. Anal Chem. 2009; 81(9):3623-9. DOI: 10.1021/ac900002u. View

2.
Walther T, Mann M . Mass spectrometry-based proteomics in cell biology. J Cell Biol. 2010; 190(4):491-500. PMC: 2928005. DOI: 10.1083/jcb.201004052. View

3.
Dettmer K, Nurnberger N, Kaspar H, Gruber M, Almstetter M, Oefner P . Metabolite extraction from adherently growing mammalian cells for metabolomics studies: optimization of harvesting and extraction protocols. Anal Bioanal Chem. 2010; 399(3):1127-39. DOI: 10.1007/s00216-010-4425-x. View

4.
Roume H, Muller E, Cordes T, Renaut J, Hiller K, Wilmes P . A biomolecular isolation framework for eco-systems biology. ISME J. 2012; 7(1):110-21. PMC: 3526178. DOI: 10.1038/ismej.2012.72. View

5.
Weckwerth W, Wenzel K, Fiehn O . Process for the integrated extraction, identification and quantification of metabolites, proteins and RNA to reveal their co-regulation in biochemical networks. Proteomics. 2004; 4(1):78-83. DOI: 10.1002/pmic.200200500. View