PMID:11309116
| Citation |
Oresnik, IJ, Ladner, CL and Turner, RJ (2001) Identification of a twin-arginine leader-binding protein. Mol. Microbiol. 40:323-31 |
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| Abstract |
The transport and targeting of a number of periplasmic proteins is carried out by the Sec-independent Mtt (also referred to as Tat) protein translocase. Proteins using this translocase have a distinct twin-arginine-containing leader. We hypothesized that specific leader-binding proteins exist to escort proteins to the translocase complex. A fusion was constructed with the twin-arginine leader from dimethyl sulphoxide (DMSO) reductase, subunit DmsA, to the N-terminus of glutathione-S-transferase. This leader fusion was bound to a glutathione affinity column through which an Escherichia coli anaerobic cell-free extract was passed. Proteins that bound to the leader were then separated and identified by N-terminal sequencing, which identified DnaK and a protein originating from the uncharacterized reading frame ynfI. This gene has been designated dmsD based on the findings presented in this paper. DmsD was purified as a His6 fusion and was shown to interact with preprotein forms of DmsA and TorA (trimethyl amine N-oxide reductase). A strain carrying a dmsD knock-out mutation showed a loss of anaerobic growth on glycerol-DMSO medium and reduced growth on glycerol-fumarate medium. This work suggests that DmsD is a twin-arginine leader-binding protein. |
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| Keywords |
Amino Acid Sequence; Arginine/metabolism; Bacterial Proteins/chemistry; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Carrier Proteins/chemistry; Carrier Proteins/genetics; Carrier Proteins/metabolism; Dimethyl Sulfoxide/metabolism; Escherichia coli/genetics; Escherichia coli/growth & development; Escherichia coli/metabolism; Escherichia coli Proteins; Iron-Sulfur Proteins; Membrane Transport Proteins; Molecular Sequence Data; Oxidoreductases/chemistry; Oxidoreductases/metabolism; Oxidoreductases, N-Demethylating/metabolism; Protein Sorting Signals; Protein Transport; Receptors, Cell Surface/chemistry; Receptors, Cell Surface/metabolism; Recombinant Fusion Proteins/metabolism; Sequence Analysis, DNA |
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