title: Two-dimensional gel electrophoresis-based proteomics of mycobacteria.
abstract: Two-dimensional gel electrophoresis (2-DE) in combination with mass spectrometry (MS) is the classic proteomics approach used to monitor the dynamics of protein abundance and posttranslational modifications in biological systems. In this chapter, we provide detailed protocols for 2-DE-based proteomics of mycobacteria. Adequate standard operating procedures for mycobacterial culture, subcellular fractionation, and selective enrichment of proteins are indispensable prerequisites for targeted proteome analyses. Therefore, we also provide approved protocols for selective and efficient extraction of cytosolic, secreted, and hydrophobic plasma membrane proteins of mycobacteria, as well as for isolation of mycobacteria from infected macrophages.
authors.affiliation: Department of Immunology, Max Planck Institute for Infection Biology, Charitéplatz 1, D-10117 Berlin, Germany. mattow@mpiib-berlin.mpg.de
authors.forename: Jens
authors.initials: J
authors.lastname: Mattow
authors.forename: Frank
authors.initials: F
authors.lastname: Siejak
authors.forename: Kristine
authors.initials: K
authors.lastname: Hagens
authors.forename: Julia
authors.initials: J
authors.lastname: Kreuzeder
authors.forename: Stefan H E
authors.initials: SH
authors.lastname: Kaufmann
authors.forename: Ulrich E
authors.initials: UE
authors.lastname: Schaible
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D008264:Macrophages / Q000382:microbiology; D009161:Mycobacterium / Q000302:isolation & purification / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D001426:Bacterial Proteins