Abstract Although the rapid progress of NMR technology has significantly expanded the range of NMR-trackable systems, preparation of NMR-suitable samples that are highly soluble and stable remains a bottleneck for studies of many biological systems. The application of solubility-enhancement tags (SETs) has been highly effective in overcoming solubility and sample stability issues and has enabled structural studies of important biological systems previously deemed unapproachable by solution NMR techniques. In this review, we provide a brief survey of the development and successful applications of the SET strategy in biomolecular NMR. We also comment on the criteria for choosing optimal SETs, such as for differently charged target proteins, and recent new developments on NMR-invisible SETs.
Content Type Journal Article
Pages 23-31
DOI 10.1007/s10858-009-9371-6
Authors
Pei Zhou, Duke University Medical Center Department of Biochemistry 242 Nanaline Duke Building, Research Drive Durham NC 27710 USA
Gerhard Wagner, Harvard Medical School Department of Biological Chemistry and Molecular Pharmacology Building C1, Room 112, 240 Longwood Avenue Boston MA 02115 USA
Improving protein solubility & long-term stability
Has anybody successfully used the following method?
A simple method for improving protein solubility and long-term stability.
Golovanov AP, Hautbergue GM, Wilson SA, Lian LY.
Department of Biomolecular Sciences, University of Manchester Institute of Science and Technology, P. O. Box 88, Manchester M60 1QD, UK. a.golovanov@umist.ac.jp
J Am Chem Soc. 2004 Jul 28;126(29):8933-9.
Straightforward, effective calibration of SPINAL-64 decoupling results in the enhancement of sensitivity and resolution of biomolecular solid-state NMR.
Straightforward, effective calibration of SPINAL-64 decoupling results in the enhancement of sensitivity and resolution of biomolecular solid-state NMR.
Straightforward, effective calibration of SPINAL-64 decoupling results in the enhancement of sensitivity and resolution of biomolecular solid-state NMR.
J Magn Reson. 2010 Dec 31;
Authors: Comellas G, Lopez JJ, Nieuwkoop AJ, Lemkau LR, Rienstra CM
We describe a simple yet highly effective optimization strategy for SPINAL-64 (1)H decoupling conditions for magic-angle spinning solid-state NMR. With...
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Recovering lost magnetization: polarization enhancement in biomolecular NMR
Recovering lost magnetization: polarization enhancement in biomolecular NMR
Abstract Experimental sensitivity remains a major drawback for the application of NMR spectroscopy to fragile and low concentrated biomolecular samples. Here we describe an efficient polarization enhancement mechanism in longitudinal-relaxation enhanced fast-pulsing triple-resonance experiments. By recovering undetectable 1H polarization originating from longitudinal relaxation during the pulse sequence, the steady-state 15N polarization becomes enhanced by up to a factor of ~5 with respect to thermal...
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Recovering lost magnetization: polarization enhancement in biomolecular NMR.
Recovering lost magnetization: polarization enhancement in biomolecular NMR.
Recovering lost magnetization: polarization enhancement in biomolecular NMR.
J Biomol NMR. 2010 Dec 30;
Authors: Favier A, Brutscher B
Experimental sensitivity remains a major drawback for the application of NMR spectroscopy to fragile and low concentrated biomolecular samples. Here we describe an efficient polarization enhancement mechanism in longitudinal-relaxation enhanced fast-pulsing triple-resonance experiments. By recovering undetectable (1)H polarization...
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[NMR paper] A straight-forward method of optimising protein solubility for NMR.
A straight-forward method of optimising protein solubility for NMR.
Related Articles A straight-forward method of optimising protein solubility for NMR.
J Biomol NMR. 2004 Nov;30(3):283-6
Authors: Howe PW
Maximising solubility is a key step in applying solution-state NMR to proteins. The 'microbatch' crystallisation screening method can be adapted to screen for protein solubility. In this approach, drops of test solutions are placed under paraffin oil in 96-well screening plates. This requires very small amounts of protein, is easy to set up...
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[NMR paper] A solubility-enhancement tag (SET) for NMR studies of poorly behaving proteins.
A solubility-enhancement tag (SET) for NMR studies of poorly behaving proteins.
Related Articles A solubility-enhancement tag (SET) for NMR studies of poorly behaving proteins.
J Biomol NMR. 2001 May;20(1):11-4
Authors: Zhou P, Lugovskoy AA, Wagner G
Protein-fusion constructs have been used with great success for enhancing expression of soluble recombinant protein and as tags for affinity purification. Unfortunately the most popular tags, such as GST and MBP, are large, which hinders direct NMR studies of the fusion proteins. Cleavage of the...
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11-19-2010 08:32 PM
Optimisation of protein solubility for NMR
A straight-forward method of optimising protein solubility for NMR.
Howe PW.
Analytical Sciences, Syngenta, Jealott's Hill Research Centre, Bracknell, Berkshire, RG42 6EY, UK.
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J Biomol NMR. 2004 Nov;30(3):283-6
Abstract:
Maximising solubility is a key step in applying solution-state NMR to proteins. The 'microbatch' crystallisation screening method can be adapted to screen for protein solubility. In this approach, drops of test solutions are placed under paraffin oil in 96-well screening plates. This requires very...
Overcoming the solubility limit with solubility-enhancement tags: successful applications in biomolecular NMR studies
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