ArticlePloS one2011
Optimized purification of a heterodimeric ABC transporter in a highly stable form amenable to 2-D crystallization.
Article in PloS one, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
22 citing papers in PubMed.
- Drug-bound outward-facing conformation of a heterodimeric ABC exporter suggests a putative mechanism of drug translocation.Nature communications · 2025Article
- The endoribonuclease Rae1 from Bacillus subtilis cleaves mRNA upstream of stalled ribosomes.Nucleic acids research · 2025Article
- Shutdown of multidrug transporterRNA (New York, N.Y.) · 2023Article
- Emerging Diversity in Lipid-Protein Interactions.Chemical reviews · 2019Review
- Functionality of membrane proteins overexpressed and purified from E. coli is highly dependent upon the strain.Scientific reports · 2019Article
- Review
- Activity of the purified plant ABC transporter NtPDR1 is stimulated by diterpenes and sesquiterpenes involved in constitutive and induced defenses.The Journal of biological chemistry · 2017Article
- Direct Spectroscopic Detection of ATP Turnover Reveals Mechanistic Divergence of ABC Exporters.Structure (London, England : 1993) · 2017Article
- Rae1/YacP, a new endoribonuclease involved in ribosome-dependent mRNA decay inThe EMBO journal · 2017Article
- High-efficient production and biophysical characterisation of nicastrin and its interaction with APPC100.Scientific reports · 2017Article
- Article
- Durable proteo-hybrid vesicles for the extended functional lifetime of membrane proteins in bionanotechnology.Chemical communications (Cambridge, England) · 2016Article
- The Heterodimeric ABC Transporter EfrCD Mediates Multidrug Efflux in Enterococcus faecalis.Antimicrobial agents and chemotherapy · 2016Article
- The multidrug ABC transporter BmrC/BmrD of Bacillus subtilis is regulated via a ribosome-mediated transcriptional attenuation mechanism.Nucleic acids research · 2014Article
- Article
- Efficient and stable reconstitution of the ABC transporter BmrA for solid-state NMR studies.Frontiers in molecular biosciences · 2014Article
- Stubborn contaminants: influence of detergents on the purity of the multidrug ABC transporter BmrA.PloS one · 2014Article
- ATPase activity of human ABCG1 is stimulated by cholesterol and sphingomyelin.Journal of lipid research · 2013Article
- Characterization of hydrophobic peptides in the presence of detergent by photoionization mass spectrometry.PloS one · 2013Article
- Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Optimized protocols for achieving high-yield expression, purification and reconstitution of membrane proteins are required to study their structure and function. We previously reported high-level expression in Escherichia coli of active BmrC and BmrD proteins from Bacillus subtilis, previously named YheI and YheH. These proteins are half-transporters which belong to the ABC (ATP-Binding Cassette) superfamily and associate in vivo to form a functional transporter able to efflux drugs. In this report, high-yield purification and functional reconstitution were achieved for the heterodimer BmrC/BmrD. In contrast to other detergents more efficient for solubilizing the transporter, dodecyl-ß-D-maltoside (DDM) maintained it in a drug-sensitive and vanadate-sensitive ATPase-competent state after purification by affinity chromatography. High amounts of pure proteins were obtained which were shown either by analytical ultracentrifugation or gel filtration to form a monodisperse heterodimer in solution, which was notably stable for more than one month at 4°C. Functional reconstitution using different lipid compositions induced an 8-fold increase of the ATPase activity (k(cat)∼5 s(-1)). We further validated that the quality of the purified BmrC/BmrD heterodimer is suitable for structural analyses, as its reconstitution at high protein densities led to the formation of 2-D crystals. Electron microscopy of negatively stained crystals allowed the calculation of a projection map at 20 Å resolution revealing that BmrC/BmrD might assemble into oligomers in a lipidic environment.
Indexed as
Identifiers
What OpenQuestion holds
Registered trials
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.