[NMR paper] (2S,4S)-5-Fluoroleucine, (2S,4R)-5-Fluoroleucine, and 5,5'-Difluoroleucine in Escherichia coli PpiB: Protein Production, (19)F NMR, and Ligand Sensing Enhanced by the gamma-Gauche Effect
(2S,4S)-5-Fluoroleucine, (2S,4R)-5-Fluoroleucine, and 5,5'-Difluoroleucine in Escherichia coli PpiB: Protein Production, (19)F NMR, and Ligand Sensing Enhanced by the gamma-Gauche Effect
Global substitution of leucine for analogues containing CH(2)F instead of methyl groups delivers proteins with multiple sites for monitoring by ^(19)F nuclear magnetic resonance (NMR) spectroscopy. The 19 kDa Escherichia coli peptidyl-prolyl cis-trans isomerase B (PpiB) was prepared with uniform high-level substitution of leucine by (2S,4S)-5-fluoroleucine, (2S,4R)-5-fluoroleucine, or 5,5'-difluoroleucine. The stability of the samples toward thermal denaturation was little altered compared to...
[NMR paper] Strategies for the production of isotopically labelled Fab fragments of therapeutic antibodies in Komagataella phaffii (Pichia pastoris) and Escherichia coli for NMR studies
Strategies for the production of isotopically labelled Fab fragments of therapeutic antibodies in Komagataella phaffii (Pichia pastoris) and Escherichia coli for NMR studies
The importance and fast growth of therapeutic monoclonal antibodies, both innovator and biosimilar products, have triggered the need for the development of characterization methods at high resolution such as nuclear magnetic resonance (NMR) spectroscopy. However, the full power of NMR spectroscopy cannot be unleashed without labelling the mAb of interest with NMR-active isotopes. Here, we present strategies using...
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12-01-2023 06:23 PM
[NMR paper] NMR resonance assignments for the nucleotide binding domains of the E. coli clamp loader complex gamma subunit
NMR resonance assignments for the nucleotide binding domains of the E. coli clamp loader complex gamma subunit
The E. coli ? clamp loader is a pentameric complex of ?, ?' and three ? subunits that opens and loads ?-clamp proteins onto DNA in an ATP-dependent process essential for efficient DNA replication. ATP binding to the ? subunits promotes conformational changes that enable the clamp loader to bind and open the ring-shaped ?-clamp homodimer. Here we report the nearly complete backbone and side-chain ¹H, ^(13)C and ^(15)N NMR resonance assignments of the 242-residue truncated ? subunit...
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03-24-2021 11:20 PM
Efficient production of a functional G protein-coupled receptor in E. coli for structural studies
Efficient production of a functional G protein-coupled receptor in E. coli for structural studies
Abstract
G protein-coupled receptors (GPCRs) are transmembrane signal transducers which regulate many key physiological process. Since their discovery, their analysis has been limited by difficulties in obtaining sufficient amounts of the receptors in high-quality, functional form from heterologous expression hosts. Albeit highly attractive because of its simplicity and the ease of isotope labeling for NMR studies, heterologous expression of functional...
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01-27-2021 04:52 AM
[NMR paper] 1H, 13C and 15*N NMR assignments of solubility tag protein Msyb of Escherichia coli.
1H, 13C and 15*N NMR assignments of solubility tag protein Msyb of Escherichia coli.
Related Articles 1H, 13C and 15*N NMR assignments of solubility tag protein Msyb of Escherichia coli.
Biomol NMR Assign. 2020 Jun 05;:
Authors: Yang Y, Ke Z, Wang Z, Li Y, Li Y, Wang Y, Liu B
Abstract
Biochemical and structural characterizations of a protein are the prerequisite for the further understanding of its biological role and potential applications. The expression of recombinant protein is almost unavoidable to produce the amount of the...
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06-08-2020 02:01 AM
High speed 3D overhauser-enhanced MRI using combined b-SSFP and compressed sensing
From The DNP-NMR Blog:
High speed 3D overhauser-enhanced MRI using combined b-SSFP and compressed sensing
Sarracanie, M., et al., High speed 3D overhauser-enhanced MRI using combined b-SSFP and compressed sensing. Magn Reson Med, 2013. 71(2): p. 735-745.
http://www.ncbi.nlm.nih.gov/pubmed/23475813
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03-24-2014 11:08 PM
[NMR paper] Strategies for Protein NMR in Escherichia coli.
Strategies for Protein NMR in Escherichia coli.
Related Articles Strategies for Protein NMR in Escherichia coli.
Biochemistry. 2014 Mar 5;
Authors: Xu G, Ye Y, Liu X, Cao S, Wu Q, Cheng K, Liu M, Pielak GJ, Li C
Abstract
In-cell NMR spectroscopy provides insight into protein conformation, dynamics and function at atomic resolution in living cells. Systematic evaluation of isotopic labeling strategies is necessary to observe the target protein in the sea of other molecules in the cell. Here, we investigate the detectability, sensitivity and...
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03-08-2014 06:52 AM
(1)H, (13)C and (15)N NMR assignments of the Escherichia coli Orf135 protein.
(1)H, (13)C and (15)N NMR assignments of the Escherichia coli Orf135 protein.
(1)H, (13)C and (15)N NMR assignments of the Escherichia coli Orf135 protein.
Biomol NMR Assign. 2011 May 7;
Authors: Kawasaki K, Yoneyama M, Murata-Kamiya N, Harashima H, Kojima C, Ito Y, Kamiya H, Mishima M
Escherichia coli Orf135 protein is thought to be an enzyme that efficiently hydrolyzes oxidatively damaged nucleotides such as 2-hydroxy-dATP, 8-hydroxy-dGTP and 5-hydroxy-CTP, in addition to 5-methyl-dCTP, dCTP and CTP, thus preventing mutations in cells caused by...
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05-10-2011 05:11 PM
[NMR paper] Identification of the DNA binding surface of H-NS protein from Escherichia coli by he
Identification of the DNA binding surface of H-NS protein from Escherichia coli by heteronuclear NMR spectroscopy.
Related Articles Identification of the DNA binding surface of H-NS protein from Escherichia coli by heteronuclear NMR spectroscopy.
FEBS Lett. 1999 Jul 16;455(1-2):63-9
Authors: Shindo H, Ohnuki A, Ginba H, Katoh E, Ueguchi C, Mizuno T, Yamazaki T
The DNA binding domain of H-NS protein was studied with various N-terminal deletion mutant proteins and identified by gel retardation assay and heteronuclear 2D- and 3D-NMR...
(2S,4S)-5-Fluoroleucine, (2S,4R)-5-Fluoroleucine, and 5,5'-Difluoroleucine in Escherichia coli PpiB: Protein Production, (19)F NMR, and Ligand Sensing Enhanced by the gamma-Gauche Effect
Linear Mode
