title: A method to investigate protein association with intact sealed mycobacterial membrane vesicles.
abstract: In mycobacteria, probing the association of cytoplasmic proteins with the membrane itself, as well as with integral or peripheral membrane proteins, is limited by the difficulty in extracting intact sealed membrane vesicles due to the complex cell wall structure. Here we tested the association of Mycobacterium tuberculosis SecA1 and SecA2 proteins with intact membrane vesicles by a flotation assay using iodixanol density gradients. These protocols have wide applications for studying the association of other mycobacterial cytoplasmic proteins with the membrane and membrane-associated proteins.
authors.affiliation: Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT 06269, USA.
authors.forename: Nadia G
authors.initials: NG
authors.lastname: D'Lima
authors.affiliation: Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT 06269, USA; Department of Chemistry, University of Connecticut, Storrs, CT 06269, USA. Electronic address: carolyn.teschke@uconn.edu.
authors.forename: Carolyn M
authors.initials: CM
authors.lastname: Teschke
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000737:chemistry* / Q000378:metabolism; D009169:Mycobacterium tuberculosis / Q000737:chemistry* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D026901:Membrane Transport Proteins; D000251:Adenosine Triphosphatases; C442122:SecA1 protein, Mycobacterium tuberculosis; C442123:SecA2 protein, Mycobacterium
keywords: Bacterial protein secretion; Membrane association; Mycobacteria