Graphene and graphene oxide as a solid matrix for extraction of membrane and membrane-associated proteins

• Published in: Mikrochimica acta (1966) Vol. 185; no. 2; pp. 123 - 10
• Main Authors: Uzzaman, Asad, Shang, Zhi, Qiao, Zhi, Cao, Cheng-Xi, Xiao, Hua
• Format: Journal Article
• Language: English
• Published: Vienna Springer Vienna 01.02.2018; Springer; Springer Nature B.V
• Subjects: Analytical Chemistry; Cancer; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Gene expression; Graphene; Graphite; Graphite - chemistry; Humans; Hydrophobic and Hydrophilic Interactions; Hydrophobicity; Immunoassay; Lipids; Membrane lipids; Membrane proteins; Membrane Proteins - isolation & purification; Membranes - chemistry; Microengineering; Nanochemistry; Nanomaterials; Nanotechnology; Neoplasm Proteins - isolation & purification; Original Paper; Proteins; Proteomics; Proteomics - methods; Solid Phase Extraction; Solid Phase Microextraction; Solubilization; Sorbents; Hydrophobic-hydrophobic interaction; Immunoassay; Cellular membrane protein; Proteomics; Preconcentration; Microextraction; Mass spectrometry; Carbon nanomaterials; Cancer
• ISSN: 0026-3672; 1436-5073; 1436-5073
• DOI: 10.1007/s00604-017-2658-5

The extraction of membrane proteins remain a challenge due to innate hydrophobicity, dynamic discrepancy, and restrain effect of membrane lipids. Nanomaterials with high surface area have competency of hydrophobic-hydrophobic lipid interactions. It is shown here that both graphene and graphene oxide dissolved in solubilization buffer are viable sorbents for efficient extraction of membrane proteins. LC-MS/MS analysis further revealed that graphene (50–200 nm) and graphene oxide (50–200 nm) can enrich more kinds of membrane proteins than a commercially available kit. Graphene was further applied to the enrichment of membrane proteins of normal cells as well as cancer cells, and 1079 and 872 proteins were identified, respectively, among which 56.5% and 60.5% were membrane proteins. In particular, 241 proteins were significantly regulated in cancer cells. Gene expression of 15 proteins was verified by qRT-PCR, and 4 of them were further quantified by immunoassay. These data collectively demonstrate that graphene has great potential to improve membrane protein extractions and thus can serve downstream cancer proteomics. Graphical abstract Two dimensional carbon nanomaterials, including graphene and graphene oxide, were employed as solid matrix to avoid lipid bilayer interference and enhance the extraction efficiency of membrane and membrane associated proteins. The strategy will benefit downstream membrane proteomics analysis.