Isolation of cell surface proteins for mass spectrometry-based proteomics

2010 ◽  
Vol 7 (1) ◽  
pp. 141-154 ◽  
Author(s):  
Sarah Elschenbroich ◽  
Yunee Kim ◽  
Jeffrey A Medin ◽  
Thomas Kislinger
2021 ◽  
Author(s):  
Anna Cioce ◽  
Beatriz Calle ◽  
Andrea Marchesi ◽  
Ganka Bineva-Todd ◽  
Helen Flynn ◽  
...  

Interactions between cells fundamentally impact biological processes. In cancer development, such interactions define key stages of disease that cannot be adequately recapitulated in cell monoculture. Complex co-culture studies have been key to unraveling the complexity of these processes, usually by sorting cells and transcriptome or bulk proteome analyses. However, these methods invariably lead to sample loss and do not capture aberrant glycosylation as an important corollary of cancer formation. Here, we report the development of Bio-Orthogonal Cell line-specific Tagging of Glycoproteins (BOCTAG). Cells are equipped with a biosynthetic AND gate that uses bioorthogonally tagged sugars to generate glycosylation precursors. The cellular glycosylation machinery then introduces bioorthogonal tags into glycoproteins exclusively in cell lines expressing the enzymes of the biosynthetic AND gate. Modification with clickable reporter moieties allows for imaging or enrichment with mass spectrometry-proteomics in a cell-specific fashion. Making use of glycans as a property of most cell surface proteins, we use BOCTAG as an efficient means for cell-specific protein tracing.


2007 ◽  
Vol 319 (1-2) ◽  
pp. 21-33 ◽  
Author(s):  
Alexander V. Filatov ◽  
Grigory I. Krotov ◽  
Victor G. Zgoda ◽  
Yuri Volkov

Author(s):  
Yunee Kim ◽  
Sarah Elschenbroich ◽  
Parveen Sharma ◽  
Lusia Sepiashvili ◽  
Anthony O. Gramolini ◽  
...  

Author(s):  
Watt W. Webb

Plasma membrane heterogeneity is implicit in the existence of specialized cell surface organelles which are necessary for cellular function; coated pits, post and pre-synaptic terminals, microvillae, caveolae, tight junctions, focal contacts and endothelial polarization are examples. The persistence of these discrete molecular aggregates depends on localized restraint of the constituent molecules within specific domaines in the cell surface by strong intermolecular bonds and/or anchorage to extended cytoskeleton. The observed plasticity of many of organelles and the dynamical modulation of domaines induced by cellular signaling evidence evanescent intermolecular interactions even in conspicuous aggregates. There is also strong evidence that universal restraints on the mobility of cell surface proteins persist virtually everywhere in cell surfaces, not only in the discrete organelles. Diffusion of cell surface proteins is slowed by several orders of magnitude relative to corresponding protein diffusion coefficients in isolated lipid membranes as has been determined by various ensemble average methods of measurement such as fluorescence photobleaching recovery(FPR).


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