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Purification of a Tat leader peptide by co-expression with its chaperone
- Source :
-
Protein Expression & Purification . Jul2012, Vol. 84 Issue 1, p167-172. 6p. - Publication Year :
- 2012
-
Abstract
- Abstract: We present a method for the purification of the 45 residue long leader peptide of Escherichia coli dimethyl sulfoxide reductase subunit A (DmsAL), a substrate of the twin arginine translocase, by co-expressing the leader peptide with its specific chaperone protein, DmsD. The peptide can be isolated from the soluble DmsAL/DmsD complex or conveniently from the lysate pellet fraction. The recombinant leader peptide is functionally intact as the peptide/chaperone complex can be reconstituted from purified DmsAL and DmsD. A construct with DmsAL fused to the N-terminus of DmsD (DmsAL–DmsD fusion) was created to further explore the properties of the leader peptide-chaperone interactions. Analytical size-exclusion chromatography in-line with multi-angle light scattering reveals that the DmsAL–DmsD fusion construct forms a dimer wherein each protomer binds the neighboring leader peptide. A model of this homodimeric interaction is presented. [Copyright &y& Elsevier]
Details
- Language :
- English
- ISSN :
- 10465928
- Volume :
- 84
- Issue :
- 1
- Database :
- Academic Search Index
- Journal :
- Protein Expression & Purification
- Publication Type :
- Academic Journal
- Accession number :
- 76610050
- Full Text :
- https://doi.org/10.1016/j.pep.2012.05.002