ArticleProtein science : a publication of the Protein Society2005
Bcl-XL as a fusion protein for the high-level expression of membrane-associated proteins.
Article in Protein science : a publication of the Protein Society, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 26 citations in OpenAlex.
- Bcl-xL Is Spontaneously Inserted into Preassembled Nanodiscs and Stimulates Bax Insertion in a Cell-Free Protein Synthesis System.Biomolecules · 2023Article
- Membrane protein structure from rotational diffusion.Biochimica et biophysica acta · 2015Review
- High-yield membrane protein expression from E. coli using an engineered outer membrane protein F fusion.Protein science : a publication of the Protein Society · 2013Article
- Expression and purification of the p75 neurotrophin receptor transmembrane domain using a ketosteroid isomerase tag.Microbial cell factories · 2012Article
- Article
- Effects of PKA phosphorylation on the conformation of the Na,K-ATPase regulatory protein FXYD1.Biochimica et biophysica acta · 2009Article
- A general strategy for the bacterial expression of amyloidogenic peptides using BCL-XL-1/2 fusions.Protein science : a publication of the Protein Society · 2009Article
- Making the most of fusion tags technology in structural characterization of membrane proteins.Molecular biotechnology · 2009Review
- Membrane orientation of the Na,K-ATPase regulatory membrane protein CHIF determined by solid-state NMR.Magnetic resonance in chemistry : MRC · 2007Article
- Structural biology of transmembrane domains: efficient production and characterization of transmembrane peptides by NMR.Protein science : a publication of the Protein Society · 2007Article
- Structure of the Na,K-ATPase regulatory protein FXYD1 in micelles.Biochemistry · 2007Article
- NMR of membrane proteins in micelles and bilayers: the FXYD family proteins.Methods (San Diego, Calif.) · 2007Article
- Nuclear magnetic resonance structural studies of membrane proteins in micelles and bilayers.Methods in molecular biology (Clifton, N.J.) · 2007Review
- Correlation of gene and protein structures in the FXYD family proteins.Journal of molecular biology · 2005Article
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
An Escherichia coli plasmid vector for the high-level expression of hydrophobic membrane proteins is described. The plasmid, pBCL, directs the expression of a target polypeptide fused to the C terminus of a mutant form of the anti-apoptotic Bcl-2 family protein, Bcl-XL, where the hydrophobic C terminus has been deleted, and Met residues have been mutated to Leu to facilitate CNBr cleavage after a single Met inserted at the beginning of the target sequence. Fusion protein expression is in inclusion bodies, simplifying the protein purification steps. Here we report the high-level production of PLM, a membrane protein that is a member of the FXYD family of tissue-specific and physiological-state-specific auxiliary subunits of the Na,K-ATPase, expressed abundantly in heart and skeletal muscle. We demonstrate that milligram quantities of pure, isotopically labeled protein can be obtained easily and in little time with this system.
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