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Measurement of Effector Protein Translocation Using Phosphorylatable Epitope Tags and Phospho-Specific Antibodies
Journal article

Measurement of Effector Protein Translocation Using Phosphorylatable Epitope Tags and Phospho-Specific Antibodies

Sara Schesser Bartra and Gregory V Plano
Methods in molecular biology (Clifton, N.J.), Vol.1531, pp.111-119
2017
PMID: 27837486

Abstract

Recombinant Fusion Proteins - immunology Amino Acid Sequence Cell Line Phosphorylation Yersinia pestis - metabolism Bacterial Proteins - chemistry Recombinant Fusion Proteins - chemistry Recombinant Fusion Proteins - metabolism Blotting, Western Protein Transport Epitopes - immunology Yersinia pestis - immunology Animals Antibodies, Phospho-Specific - immunology Plasmids - genetics Bacterial Proteins - metabolism Bacterial Secretion Systems - metabolism Epitopes - chemistry Mice Gene Order
Numerous bacterial pathogens employ specialized protein secretion machineries to directly inject anti-host proteins, termed effector proteins, into eukaryotic cells. Effector proteins carrying small phosphorylatable tags can be used to detect and quantify effector protein injection. Here, we describe the use of the ELK- and GSK-tags to detect the translocation of the Y. pestis YopE effector protein into RAW 264.7 macrophage-like cells using immunoblot analysis with phospho-specific antibodies.

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