Altered Escherichia coli membrane protein assembly machinery allows proper membrane assembly of eukaryotic protein vitamin K epoxide reductase
| Autor: | Yongxin Zhao, Eric Mandela, Jonathan Beckwith, Dana Boyd, Robert E. Campbell, Feras Hatahet, Jessica L. Blazyk, Eugénie Martineau |
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| Rok vydání: | 2015 |
| Předmět: |
chemistry.chemical_classification
Mutation Vesicle-associated membrane protein 8 Multidisciplinary Escherichia coli Proteins Mutant Membrane Proteins Biology Chromosomes Bacterial Biological Sciences medicine.disease_cause Rats Membrane Enzyme Membrane protein Biochemistry chemistry Vitamin K Epoxide Reductases medicine Escherichia coli Animals Vitamin K epoxide reductase |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America. 112(49) |
| ISSN: | 1091-6490 |
| Popis: | Significance Proteins embedded in biological membranes are important targets for therapeutic intervention. However, they tend to be far more challenging to deal with than soluble proteins. We have used the anticoagulant target VKORc1 to investigate membrane protein expression bottlenecks in the bacterium Escherichia coli . When this poorly expressed enzyme is mutagenized or introduced to a library of E. coli mutants, functional expression is achieved. This approach gave clues on why certain foreign membrane proteins fail to correctly assimilate in E. coli membranes including mis-matches in membrane protein assembly signals. We also identified key critical residues in two components of membrane protein assembly and quality control for rational manipulations to enhance functional expression. Finally, we demonstrate the high-throughput screening for potential anticoagulants using bacteria. |
| Databáze: | OpenAIRE |
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