ArticleProtein expression and purification2008
Wheat germ cell-free translation, purification, and assembly of a functional human stearoyl-CoA desaturase complex.
Article in Protein expression and purification, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 40 papers.
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Who cites it
40 citing papers in PubMed, 92 citations in OpenAlex.
- Eukaryotic cell-free protein synthesis: Chassis diversification, system engineering, and emerging applications.Synthetic and systems biotechnology · 2026Review
- Cell-free protein synthesis and vesicle systems for programmable therapeutic manufacturing and delivery.Journal of biological engineering · 2025Review
- Half a Century of Progress: The Evolution of Wheat Germ-Based In Vitro Translation into a Versatile Protein Production Method.International journal of molecular sciences · 2025Review
- Identification and functional prediction of lncRNAs associated with intramuscular lipid deposition in Guangling donkeys.Frontiers in veterinary science · 2024Article
- RNA Sequencing (RNA-Seq) Analysis Reveals Liver Lipid Metabolism Divergent Adaptive Response to Low- and High-Salinity Stress in Spotted Scat (Animals : an open access journal from MDPI · 2023Article
- RNA-Seq Study on theAnimals : an open access journal from MDPI · 2023Article
- Evaluation of Cell-Free Synthesized Human Channel Proteins for In Vitro Channel Research.Membranes · 2022Article
- Alterations of Lipid Profile in Livers with Impaired Lipophagy.International journal of molecular sciences · 2022Article
- Review
- Cell-Free Expression of a Plant Membrane Protein BrPT2 From Boesenbergia Rotunda.Molecular biotechnology · 2021Article
- Easy Synthesis of Complex Biomolecular Assemblies: Wheat Germ Cell-Free Protein Expression in Structural Biology.Frontiers in molecular biosciences · 2021Review
- Structure and Mechanism of a Unique Diiron Center in Mammalian Stearoyl-CoA Desaturase.Journal of molecular biology · 2020Article
- Metabolic Plasticity in Ovarian Cancer Stem Cells.Cancers · 2020Article
- A User's Guide to Cell-Free Protein Synthesis.Methods and protocols · 2019Review
- LRET Determination of Molecular Distances during pH Gating of the Mammalian Inward Rectifier Kir1.1b.Biophysical journal · 2018Article
- Substrate specificity and membrane topologies of the iron-containing ω3 and ω6 desaturases from Mortierella alpina.Applied microbiology and biotechnology · 2018Article
- Article
- Compartmentalization and Transport in Synthetic Vesicles.Frontiers in bioengineering and biotechnology · 2016Review
- Comprehensive study of liposome-assisted synthesis of membrane proteins using a reconstituted cell-free translation system.Scientific reports · 2015Article
- Cell-Free Protein Synthesis: Pros and Cons of Prokaryotic and Eukaryotic Systems.Chembiochem : a European journal of chemical biology · 2015Review
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
A wheat germ cell-free extract was used to perform in vitro translation of human stearoyl-CoA desaturase in the presence of unilamelar liposomes, and near complete transfer of the expressed integral membrane protein into the liposome was observed. Moreover, co-translation of the desaturase along with human cytochrome b(5) led to transfer of both membrane proteins into the liposomes. A simple, single step purification via centrifugation in a density gradient yielded proteoliposomes with the desaturase in high purity as judged by capillary electrophoresis. After in vitro reconstitution of the non-heme iron and heme active sites, the function of the reconstituted enzyme complex was demonstrated by conversion of stearoyl-CoA to oleoyl-CoA. This simple translation approach obviates the use of detergents or other lipids to stabilize and isolate a catalytically active integral membrane enzyme. The applicability of cell-free translation to the assembly and purification of other integral membrane protein complexes is discussed.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.