PMID:14976313
| Citation |
Masip, L, Pan, JL, Haldar, S, Penner-Hahn, JE, DeLisa, MP, Georgiou, G, Bardwell, JC and Collet, JF (2004) An engineered pathway for the formation of protein disulfide bonds. Science 303:1185-9 |
|---|---|
| Abstract |
We have engineered a pathway for the formation of disulfide bonds. By imposing evolutionary pressure, we isolated mutations that changed thioredoxin, which is a monomeric disulfide reductase, into a [2Fe-2S] bridged dimer capable of catalyzing O2-dependent sulfhydryl oxidation in vitro. Expression of the mutant protein in Escherichia coli with oxidizing cytoplasm and secretion via the Tat pathway restored disulfide bond formation in strains that lacked the complete periplasmic oxidative machinery (DsbA and DsbB). The evolution of [2Fe-2S] thioredoxin illustrates how mutations within an existing scaffold can add a cofactor and markedly change protein function. |
| Links |
PubMed Online version:10.1126/science.1092612 |
| Keywords |
Amino Acid Motifs; Amino Acid Sequence; Amino Acid Substitution; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Cell Membrane/metabolism; Cysteine/analysis; Cytoplasm/metabolism; Dimerization; Directed Molecular Evolution; Disulfides/chemistry; Disulfides/metabolism; Escherichia coli/genetics; Escherichia coli/metabolism; Escherichia coli/physiology; Hirudins/chemistry; Hirudins/metabolism; Iron/analysis; Membrane Proteins/genetics; Membrane Proteins/metabolism; Movement; Mutation; Oxidation-Reduction; Oxygen/metabolism; Protein Disulfide-Isomerases/genetics; Protein Disulfide-Isomerases/metabolism; Protein Engineering; Protein Folding; Proteins/chemistry; Proteins/metabolism; Sulfides/analysis; Thioredoxins/chemistry; Thioredoxins/genetics; Thioredoxins/metabolism |
| edit table |
Significance
You can help EcoliWiki by summarizing why this paper is useful
Useful Materials and Methods
You can help Ecoliwiki by describing the useful materials (strains, plasmids, antibodies, etc) described in this paper.
Annotations
<annotationlinks/>
EcoliWiki Links
Add links to pages that link here (e.g. gene, product, method pages)
References
See Help:References for how to manage references in EcoliWiki.