Solution NMR studies of large proteins are hampered by rapid signal decay due to short-range dipolar ¹H-¹H and ¹H-^(13)C interactions. These are attenuated by rapid rotation in methyl groups and by deuteration (²H), so selective ¹H,^(13)C-isotope labelling of methyl groups in otherwise perdeuterated proteins, combined with methyl transverse relaxation optimized spectroscopy (methyl-TROSY), is now standard for solution NMR of large protein systems > 25 kDa. For non-methyl positions, long-lived...
[NMR paper] Cost-effective large-scale expression of proteins for NMR studies.
Cost-effective large-scale expression of proteins for NMR studies.
Related Articles Cost-effective large-scale expression of proteins for NMR studies.
J Biomol NMR. 2018 May 19;:
Authors: Klopp J, Winterhalter A, Gébleux R, Scherer-Becker D, Ostermeier C, Gossert AD
Abstract
We present protocols for high-level expression of isotope-labelled proteins in E. coli in cost-effective ways. This includes production of large amounts of unlabeled proteins and 13C-methyl methionine labeling in rich media, where yields of up to a gram of...
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05-21-2018 06:16 PM
Cost-effective large-scale expression of proteins for NMR studies
Cost-effective large-scale expression of proteins for NMR studies
Abstract
We present protocols for high-level expression of isotope-labelled proteins in E. coli in cost-effective ways. This includes production of large amounts of unlabeled proteins and 13C-methyl methionine labeling in rich media, where yields of up to a gram of soluble protein per liter of culture are reached. Procedures for uniform isotope labeling of 2H, 13C and 15N using auto-induction or isopropyl-β-d-1-thiogalactopyranoside-induction are described, with primary focus on...
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05-19-2018 05:41 PM
Storage of nuclear magnetization as long-lived singlet order in low magnetic field
From The DNP-NMR Blog:
Storage of nuclear magnetization as long-lived singlet order in low magnetic field
This week I will catch up with some articles that were published in the Proceedings of the National Academy of Sciences that slipped through the crack.
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07-21-2014 05:32 PM
A cost-effective look at proteins - with a little help from robotics - Cordis News
http://www.bionmr.com//t3.gstatic.com/images?q=tbn:ANd9GcTypMhbcsoTBmkGzm1ZmzjTHj-b-AsNVL4y8L03Y0oBfj2cCZUDv2gZmGEzmim-0VUKjKStcwU
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A cost-effective look at proteins - with a little help from robotics
Cordis News
Until now, a number of methods have been used to study static protein structures, including X-ray crystallography and nuclear magnetic resonance. However, these methods are of no use for proteins that are on the move; analytical methods, computer ...
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A cost-effective look at proteins - with a little help from...
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09-14-2013 01:46 AM
Magnetic Resonance Force Microscopy Detected Long-Lived Spin Magnetization
From The DNP-NMR Blog:
Magnetic Resonance Force Microscopy Detected Long-Lived Spin Magnetization
Chen, L., et al., Magnetic Resonance Force Microscopy Detected Long-Lived Spin Magnetization. Magnetics, IEEE Transactions on, 2013. 49(7): p. 3528-3532.
http://dx.doi.org/10.1109/TMAG.2013.2239268
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08-23-2013 02:29 PM
Amino acid selective unlabeling for sequence specific resonance assignments in proteins
Amino acid selective unlabeling for sequence specific resonance assignments in proteins
Abstract Sequence specific resonance assignment constitutes an important step towards high-resolution structure determination of proteins by NMR and is aided by selective identification and assignment of amino acid types. The traditional approach to selective labeling yields only the chemical shifts of the particular amino acid being selected and does not help in establishing a link between adjacent residues along the polypeptide chain, which is important for sequential assignments. An alternative...
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03-20-2012 12:42 AM
4D APSY-HBCB(CG)CDHD experiment for automated assignment of aromatic amino acid side chains in proteins
4D APSY-HBCB(CG)CDHD experiment for automated assignment of aromatic amino acid side chains in proteins
Abstract A four-dimensional (4D) APSY (automated projection spectroscopy)-HBCB(CG)CDHD experiment is presented. This 4D experiment correlates aromatic with aliphatic carbon and proton resonances from the same amino acid side chain of proteins in aqueous solution. It thus allows unambiguous sequence-specific assignment of aromatic amino acid ring signals based on backbone assignments. Compared to conventional 2D approaches, the inclusion of evolution periods on 1Hβ and 13Cδ...
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09-30-2011 08:01 PM
[NMR paper] Cell-free synthesis and amino acid-selective stable isotope labeling of proteins for
Cell-free synthesis and amino acid-selective stable isotope labeling of proteins for NMR analysis.
Related Articles Cell-free synthesis and amino acid-selective stable isotope labeling of proteins for NMR analysis.
J Biomol NMR. 1995 Sep;6(2):129-34
Authors: Kigawa T, Muto Y, Yokoyama S
For the application of multidimensional NMR spectroscopy to larger proteins, it would be useful to perform selective labeling of one of the 20 amino acids. For some amino acids, however, amino acid metabolism drastically reduces the efficiency and selectivity...