ArticleBMC structural biology2004
Fast identification of folded human protein domains expressed in E. coli suitable for structural analysis.
Article in BMC structural biology, 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 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
9 citing papers in PubMed.
- An Evolution-Based Approach to De Novo Protein Design.Methods in molecular biology (Clifton, N.J.) · 2017Article
- Article
- Identification of soluble protein fragments by gene fragmentation and genetic selection.Nucleic acids research · 2008Article
- Small-scale, semi-automated purification of eukaryotic proteins for structure determination.Journal of structural and functional genomics · 2007Article
- Effect of N-terminal solubility enhancing fusion proteins on yield of purified target protein.Journal of structural and functional genomics · 2006Article
- Towards miniaturization of a structural genomics pipeline using micro-expression and microcoil NMR.Journal of structural and functional genomics · 2005Article
- Structural genomics of human proteins--target selection and generation of a public catalogue of expression clones.Microbial cell factories · 2005Article
- Production of soluble mammalian proteins in Escherichia coli: identification of protein features that correlate with successful expression.BMC biotechnology · 2004Article
- Review
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
9 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundHigh-throughput protein structure analysis of individual protein domains requires analysis of large numbers of expression clones to identify suitable constructs for structure determination. For this purpose, methods need to be implemented for fast and reliable screening of the expressed proteins as early as possible in the overall process from cloning to structure determination.
results88 different E. coli expression constructs for 17 human protein domains were analysed using high-throughput cloning, purification and folding analysis to obtain candidates suitable for structural analysis. After 96 deep-well microplate expression and automated protein purification, protein domains were directly analysed using 1D 1H-NMR spectroscopy. In addition, analytical hydrophobic interaction chromatography (HIC) was used to detect natively folded protein. With these two analytical methods, six constructs (representing two domains) were quickly identified as being well folded and suitable for structural analysis.
conclusionThe described approach facilitates high-throughput structural analysis. Clones expressing natively folded proteins suitable for NMR structure determination were quickly identified upon small scale expression screening using 1D 1H-NMR and/or analytical HIC. This procedure is especially effective as a fast and inexpensive screen for the 'low hanging fruits' in structural genomics.
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.