Isolation of cell surface proteins for mass spectrometry-based proteomics

Abstract

Defining the cell surface proteome has profound importance for understanding cell differentiation and cell–cell interactions, as well as numerous pathogenic abnormalities. Owing to their hydrophobic nature, plasma membrane proteins that reside on the cell surface pose analytical challenges and, despite efforts to overcome difficulties, remain under-represented in proteomic studies. Limitations in the classically employed ultracentrifugation-based approaches have led to the invention of more elaborate techniques for the purification of cell surface proteins. Three of these methods – cell surface coating with cationic colloidal silica beads, biotinylation and chemical capture of surface glycoproteins – allow for marked enrichment of this subcellular proteome, with each approach offering unique advantages and characteristics for different experiments. In this article, we introduce the principles of each purification method and discuss applications from the recent literature.

Keywords::

• biotinylation
• cell surface
• cell surface capture
• mass spectrometry
• plasma membrane
• proteomics
• silica-bead coating

Financial & competing interests disclosure

Thomas Kislinger is supported through the Canadian Research Chairs Program. This work was supported in parts by a grant from the Canadian Institute Health Research (MOP-93772) to Thomas Kislinger and Jeffrey Medin. Yunee Kim is a recipient of a Paul Starita Graduate Student Fellowships. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.