In vitro analysis of the bacterial twin-arginine-dependent protein export.
Prokaryotic organisms possess a specialized protein translocase in their cytoplasmic membranes that catalyzes the export of folded preproteins. Substrates for this pathway are distinguished by a twin-arginine consensus motif in their signal peptides (twin-arginine translocation [Tat] pathway). We have compiled detailed protocols for the preparation and operation of a cell-free system by which the bacterial Tat pathway can be fully reproduced in vitro. This system has proven useful and is being further exploited for the study of precursor-translocase interactions, assembly of the translocase, and the mechanism of transmembrane passage.
Moser
Michael
M
Institut für Biochemie und Molekularbiologie, Zentrum fiir Biochemie und Molekulare Zellforschung, Universität Freiburg, Freiburg, Germany.
Panahandeh
Sascha
S
Holzapfel
Eva
E
Müller
Matthias
M
D001426:Bacterial Proteins / Q000032:analysis* / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002474:Cell-Free System; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D066298:In Vitro Techniques; D026901:Membrane Transport Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D009492:Neurospora crassa / Q000378:metabolism; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000378:metabolism
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins; C408585:twin-arginine translocase complex, E coli