title: Measurement of Effector Protein Translocation Using Phosphorylatable Epitope Tags and Phospho-Specific Antibodies.
abstract: 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.
authors.affiliation: Department of Microbiology and Immunology, Miller School of Medicine, University of Miami, 1600 NW 10th Ave., Miami, FL, 33136, USA.
authors.forename: Sara Schesser
authors.initials: SS
authors.lastname: Bartra
authors.affiliation: Department of Microbiology and Immunology, Miller School of Medicine, University of Miami, 1600 NW 10th Ave., Miami, FL, 33136, USA. gplano@med.miami.edu.
authors.forename: Gregory V
authors.initials: GV
authors.lastname: Plano
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D045163:Antibodies, Phospho-Specific / Q000276:immunology; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D058947:Bacterial Secretion Systems / Q000378:metabolism*; D015153:Blotting, Western; D002460:Cell Line; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D023061:Gene Order; D051379:Mice; D010766:Phosphorylation; D010957:Plasmids / Q000235:genetics; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D015010:Yersinia pestis / Q000276:immunology / Q000378:metabolism
chemical_list: D045163:Antibodies, Phospho-Specific; D001426:Bacterial Proteins; D058947:Bacterial Secretion Systems; D000939:Epitopes; D011993:Recombinant Fusion Proteins
keywords: Effector proteins; Phospho-specific antibodies; Translocation; Type III secretion; Type IV secretion; Yersinia pestis