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

Wei Wei; Nicholas M. Riley; Xuchao Lyu; Carolyn R. Bertozzi; Jonathan Z. Long

STAR Protocols (Dec 2021)

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

Wei Wei,
Nicholas M. Riley,
Xuchao Lyu,
Carolyn R. Bertozzi,
Jonathan Z. Long

Affiliations

Wei Wei: Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Biology, Stanford University, Stanford, CA 94305, USA; Stanford ChEM-H, Stanford University, Stanford, CA 94305, USA; Corresponding author
Nicholas M. Riley: Department of Chemistry, Stanford University, Stanford, CA 94305, USA; Stanford ChEM-H, Stanford University, Stanford, CA 94305, USA; Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305, USA; Corresponding author
Xuchao Lyu: Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA; Stanford ChEM-H, Stanford University, Stanford, CA 94305, USA
Carolyn R. Bertozzi: Department of Chemistry, Stanford University, Stanford, CA 94305, USA; Stanford ChEM-H, Stanford University, Stanford, CA 94305, USA; Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305, USA
Jonathan Z. Long: Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA; Stanford ChEM-H, Stanford University, Stanford, CA 94305, USA; Corresponding author

Journal volume & issue: Vol. 2, no. 4
p. 101014

Abstract

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Summary: 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).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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