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Cell Surface Biotinylation via Furan Crosslinking to Profile the Surfaceome

ABSTRACT: The cellular surfaceome is crucial for cellular homeostasis and communication, with surface proteins acting as key receptors, transporters, and enzymes. However, comprehensive analysis of the surfaceome is challenging due to the hydrophobic nature and low abundance of membrane proteins, resulting underrepresented in mass spectrometry-based proteomic analyses. This study introduces a novel method for profiling the cell surfaceome using furan-biotin (FB) conjugates. We synthesized six FB compounds (FB1-FB6) and showed that FB1 reacts with lysine and cysteine in close proximity. FB1 exhibited the highest staining intensity and specificity for surface proteins upon oxidation. Protein pull-down assays confirmed the efficiency and reproducibility of FB1 in enriching surface proteins, outperforming traditional NHS-biotin methods by reducing nonspecific labeling of intracellular proteins. This method provides a robust tool for unbiased, high-specificity proteomic studies of cell surface dynamics, particularly under oxidative conditions, and opens new avenues for exploring cell surface interactions and protein modifications.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell

SUBMITTER: Esperanza Fernandez

LAB HEAD: Kris Gevaert

PROVIDER: PXD059552 | Pride | 2025-10-21

REPOSITORIES: Pride

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			Action	DRS
	F003371_1p_uPAC13_CMB-736_1.index	Other
	F003371_1p_uPAC13_CMB-736_1.raw	Raw
	F003373_1p_uPAC13_CMB-736_2.index	Other
	F003373_1p_uPAC13_CMB-736_2.raw	Raw
	F003375_1p_uPAC13_CMB-736_3.index	Other

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