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Protocol for cell type-specific labeling, enrichment, and proteomic profiling of plasma proteins in mice.

ABSTRACT: Secreted polypeptides represent a fundamental axis of intercellular communication. Here, we present a protocol for the cell type-specific biotinylation, enrichment, and proteomic profiling of secreted plasma proteins directly in mice. This protocol uses conditional "turn-on" adeno-associated viruses expressing an endoplasmic reticulum-targeted biotin ligase to globally biotinylate proteins of the secretory pathway in a cell type-specific manner. Biotinylated secreted proteins can be directly purified from blood plasma and analyzed by SDS-PAGE gel or shotgun proteomics. For complete information on the generation and use of this protocol, please refer to Wei et al. (2021).

SUBMITTER: Wei W

PROVIDER: S-EPMC8672099 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature

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Protocol for cell type-specific labeling, enrichment, and proteomic profiling of plasma proteins in mice.

Wei Wei W Riley Nicholas M NM Lyu Xuchao X Bertozzi Carolyn R CR Long Jonathan Z JZ

STAR protocols 20211211 4

Secreted polypeptides represent a fundamental axis of intercellular communication. Here, we present a protocol for the cell type-specific biotinylation, enrichment, and proteomic profiling of secreted plasma proteins directly in mice. This protocol uses conditional "turn-on" adeno-associated viruses expressing an endoplasmic reticulum-targeted biotin ligase to globally biotinylate proteins of the secretory pathway in a cell type-specific manner. Biotinylated secreted proteins can be directly pur ...[more]

PMID: 34950890

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