Bioorthogonal Conjugation-Assisted Purification Method for Profiling Cell Surface Proteome
Research output: Journal Publications and Reviews › RGC 21 - Publication in refereed journal › peer-review
Author(s)
Related Research Unit(s)
Detail(s)
Original language | English |
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Pages (from-to) | 1901–1909 |
Journal / Publication | Analytical Chemistry |
Volume | 94 |
Issue number | 3 |
Online published | 12 Jan 2022 |
Publication status | Published - 25 Jan 2022 |
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Abstract
Surface biotinylation has been widely adapted in profiling the cellular proteome associated with the plasma membrane. However, the workflow is subject to interference from the cytoplasmic biotin-associated proteins that compete for streptavidin-binding during purification. Here we established a that utilizes the Staudinger chemoselective ligation to label and isolate surface-associated proteins while minimizing the binding of endogenous biotin-associated proteins. Label-free quantitative proteomics demonstrated that BCAP is efficient in isolating cell surface proteins with excellent reproducibility. Subsequently, we applied BCAP to compare the surface proteome of proliferating and senescent mouse embryonic fibroblasts (MEFs). Among the results, EHD2 was identified and validated as a novel protein that is enhanced at the cell surface of senescent MEFs. We expect that BCAP will have broad applications in profiling cell surface proteomes in the future.
Research Area(s)
- SENESCENCE, IDENTIFICATION, PROTEINS
Citation Format(s)
Bioorthogonal Conjugation-Assisted Purification Method for Profiling Cell Surface Proteome. / Liu, Guopan; Choi, Ming Ho; Ma, Haiying et al.
In: Analytical Chemistry, Vol. 94, No. 3, 25.01.2022, p. 1901–1909.
In: Analytical Chemistry, Vol. 94, No. 3, 25.01.2022, p. 1901–1909.
Research output: Journal Publications and Reviews › RGC 21 - Publication in refereed journal › peer-review