Dataset Information

Proximal Biotinylation Based Combinatory Approach for Isolating Plasma Membrane Proteins

ABSTRACT: Comprehensive profiling of the cell surface proteome has been challenging due to the lack of tools for an effective and reproducible way to isolate plasma membrane proteins from mammalian cells. Here, we employ a proximity-dependent biotinylation approach to label and isolate plasma membrane proteins without an extra in vitro labeling step that we called plasma membrane-BioID. The lipid modified BirA* enzyme (MyrPalm BirA*) was targeted to the plasma membrane where it effectively biotinylates plasma membrane proteins. Biotinylated proteins were then affinity purified and analyzed by mass spectrometry. Our analysis demonstrates that combining conventional sucrose density gradient centrifugation and plasma membrane-BioID is ideal to overcome the inherent limitations of identification of integral membrane proteins and yields highly pure plasma components for downstream proteomic analysis.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Altuğ Kamacıoğlu

LAB HEAD: Nurhan Ozlu

PROVIDER: PXD018383 | Pride | 2020-08-28

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	BA_MA_20190318_OnBeadDig_MycPalm-C2-Biotin_1.raw	Raw
	BA_MA_20190318_OnBeadDig_MycPalm-C2-Biotin_2.raw	Raw
	BA_MA_20190318_OnBeadDig_MycPalm-C2_1.raw	Raw
	BA_MA_20190318_OnBeadDig_MycPalm-C2_2.raw	Raw
	BA_MA_20190426_OnBeadDig_MycPalmBiotin_1.raw	Raw

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Proximal Biotinylation-Based Combinatory Approach for Isolating Integral Plasma Membrane Proteins.

Akdag Mehmet M Yunt Zeynep Sabahat ZS Kamacioglu Altug A Qureshi Mohammed Haroon MH Akarlar Busra A BA Ozlu Nurhan N

Journal of proteome research 20200615 8

Comprehensive profiling of the cell-surface proteome has been challenging due to the lack of tools for an effective and reproducible way to isolate plasma membrane proteins from mammalian cells. Here we employ a proximity-dependent biotinylation approach to label and isolate plasma membrane proteins without an extra <i>in vitro</i> labeling step, which we call Plasma Membrane-BioID. The lipid-modified BirA* enzyme (MyrPalm BirA*) was targeted to the inner leaflet of the plasma membrane, where it ...[more]

PMID: 32500712

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