An engineered pathway for the formation of protein disulfide bonds
| Autor: | Lluis Masip, Jonathan L. Pan, George Georgiou, James E. Penner-Hahn, Jean-François Collet, James C.A. Bardwell, Matthew P. DeLisa, Suranjana Haldar |
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| Rok vydání: | 2004 |
| Předmět: |
Cytoplasm
Protein Folding Iron Movement Amino Acid Motifs Protein Disulfide-Isomerases Sulfides Protein Engineering Thioredoxins Bacterial Proteins Mutant protein Escherichia coli Amino Acid Sequence Cysteine Disulfides Protein disulfide-isomerase Multidisciplinary biology Cell Membrane Membrane Proteins Proteins Ferredoxin-thioredoxin reductase Protein engineering Hirudins Oxygen DsbA Biochemistry Amino Acid Substitution Mutation biology.protein Protein folding Thioredoxin Directed Molecular Evolution Dimerization Oxidation-Reduction |
| Zdroj: | Science (New York, N.Y.). 303(5661) |
| ISSN: | 1095-9203 |
| Popis: | 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 O 2 -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. |
| Databáze: | OpenAIRE |
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