Proteomics

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Cell Surface Engineering Enables Surfaceome Profiling

ABSTRACT: Understanding which proteins are presented at the cell surface is essential for ongoing therapeutic development, and to expand our basic understanding of biology. We envisioned the integration of cell surface engineering with radical-mediated protein biotinylation to profile cell surface proteins (CSPs). This method relies on the pre-functionalization of cells with cholesterol lipid groups, followed by their conjugation with an APEX2 ascorbate peroxidase enzyme. The APEX2 enzyme allows for the covalent conjugation of biotin moieties to nearby residues for subsequent enrichment, and serves as a tool for the enrichment and subsequent MS analysis of CSPs.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human) Mus Musculus (mouse)

TISSUE(S): Skeletal Muscle Cell, Epithelial Cell

DISEASE(S): Pancreas Adenocarcinoma,Placental Choriocarcinoma

SUBMITTER: Zak Vilen  

LAB HEAD: Mia Lace Huang

PROVIDER: PXD031845 | Pride | 2022-05-06

REPOSITORIES: Pride

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MS221059_MH_AR_ZV_B2_095.msf Msf
MS221059_MH_AR_ZV_B2_095.raw Raw
MS221086_MH_AR_AER_B2_4.msf Msf
MS221086_MH_AR_AER_B2_4.raw Raw
MS221094_MH_ZV_B2_130new.msf Msf
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Cell Surface Engineering Enables Surfaceome Profiling.

Vilen Zak Z   Reeves Abigail E AE   O'Leary Timothy R TR   Joeh Eugene E   Kamasawa Naomi N   Huang Mia L ML  

ACS chemical biology 20220420 4

Cell surface proteins (CSPs) are vital molecular mediators for cells and their extracellular environment. Thus, understanding which CSPs are displayed on cells, especially in different cell states, remains an important endeavor in cell biology. Here, we describe the integration of cell surface engineering with radical-mediated protein biotinylation to profile CSPs. This method relies on the prefunctionalization of cells with cholesterol lipid groups, followed by sortase-catalyzed conjugation wit  ...[more]

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