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All atom insights into the impact of crowded environments on protein stability by NMR spectroscopy - Nature.com
Mar 30, 2024 - 9:22 AM - by nmrlearner
nmrlearner's Avatar All atom insights into the impact of crowded environments on protein stability by NMR spectroscopy - Nature.com

All atom insights into the impact of crowded environments on protein stability by NMR spectroscopy Nature.com Read here
0 Replies | 85 Views
Temporal and spatial characterisation of protein liquid-liquid phase separation using NMR spectroscopy - Nature.com
Mar 30, 2024 - 9:22 AM - by nmrlearner
nmrlearner's Avatar Temporal and spatial characterisation of protein liquid-liquid phase separation using NMR spectroscopy - Nature.com

Temporal and spatial characterisation of protein liquid-liquid phase separation using NMR spectroscopy Nature.com Read here
0 Replies | 71 Views
Protein structure determination in living cells by in-cell NMR spectroscopy - Nature.com
Mar 30, 2024 - 9:22 AM - by nmrlearner
nmrlearner's Avatar Protein structure determination in living cells by in-cell NMR spectroscopy - Nature.com

Protein structure determination in living cells by in-cell NMR spectroscopy Nature.com Read here
0 Replies | 75 Views
Expanding the membrane-protein NMR toolkit - Nature.com
Mar 30, 2024 - 9:22 AM - by nmrlearner
nmrlearner's Avatar Expanding the membrane-protein NMR toolkit - Nature.com

Expanding the membrane-protein NMR toolkit Nature.com Read here
0 Replies | 84 Views
[NMR paper] Dimerization of the beta-Hairpin Membrane-Active Cationic Antimicrobial Peptide Capitellacin from Marine Polychaeta: An NMR Structural and Thermodynamic Study
Mar 29, 2024 - 7:50 PM - by nmrlearner
nmrlearner's Avatar Dimerization of the beta-Hairpin Membrane-Active Cationic Antimicrobial Peptide Capitellacin from Marine Polychaeta: An NMR Structural and Thermodynamic Study

Capitellacin is the ?-hairpin membrane-active cationic antimicrobial peptide from the marine polychaeta Capitella teleta. Capitellacin exhibits antibacterial activity, including against drug-resistant strains. To gain insight into the mechanism of capitellacin action, we investigated the structure of the peptide in the membrane-mimicking environment of dodecylphosphocholine (DPC) micelles using high-resolution NMR spectroscopy. In DPC solution, two structural forms of capitellacin were observed:...

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0 Replies | 166 Views
[NMR paper] Elucidation of the mechanisms of inter-domain coupling in the monomeric state of Enzyme I by high-pressure NMR
Mar 29, 2024 - 7:43 AM - by nmrlearner
nmrlearner's Avatar Elucidation of the mechanisms of inter-domain coupling in the monomeric state of Enzyme I by high-pressure NMR

The catalytic cycle of Enzyme I (EI), a phosphotransferase enzyme responsible for converting phosphoenolpyruvate (PEP) into pyruvate, is characterized by a series of local and global conformational rearrangements. This multistep process includes a monomer-to-dimer transition, followed by an open-to-closed rearrangement of the dimeric complex upon PEP binding. In the present study, we investigate the thermodynamics of EI dimerization using a range of high-pressure solution NMR techniques...

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0 Replies | 98 Views
[NMR paper] Metabolomic response to acute resistance exercise in healthy older adults by 1H-NMR
Mar 29, 2024 - 7:43 AM - by nmrlearner
nmrlearner's Avatar Metabolomic response to acute resistance exercise in healthy older adults by 1H-NMR

CONCLUSIONS: This study demonstrates that while many exercise-responsive metabolites change similarly in young and older adults, several demonstrate age-dependent changes even in the absence of evidence of sarcopenia or frailty.

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0 Replies | 89 Views
Enabling site-specific NMR investigations of therapeutic FabĀ*using a cell-free based isotopic labeling approach:Ā*application to anti-LAMP1 Fab
Mar 28, 2024 - 7:10 PM - by nmrlearner
nmrlearner's Avatar Enabling site-specific NMR investigations of therapeutic FabĀ*using a cell-free based isotopic labeling approach:Ā*application to anti-LAMP1 Fab

Abstract

Monoclonal antibodies (mAbs) are biotherapeutics that have achieved outstanding success in treating many life-threatening and chronic diseases. The recognition of an antigen is mediated by the fragment antigen binding (Fab) regions composed by four different disulfide bridge-linked immunoglobulin domains. NMR is a powerful method to assess the integrity, the structure and interaction of Fabs, but site specific analysis has been so far hampered by the size of the Fabs and the lack of approaches to produce isotopically labeled samples. We proposed here an efficient in vitro method to produce [15N, 13C, 2H]-labeled Fabs enabling high resolution NMR investigations of these powerful therapeutics. As an open system, the cell-free expression mode enables fine-tuned control of the redox potential in presence of disulfide bond isomerase to enhance the formation of native disulfide bonds. Moreover, inhibition of transaminases in the S30 cell-free extract offers the opportunity to produce perdeuterated Fab samples directly in 1H2O medium, without the need for a time-consuming and inefficient refolding process. This specific protocol was applied to produce an optimally labeled sample of a therapeutic Fab, enabling the sequential assignment of 1HN, 15N, 13Cā?², 13CĪ±, 13CĪ² resonances of a full-length Fab. 90% of the backbone resonances of a Fab domain directed against the human LAMP1 glycoprotein were assigned successfully, opening new opportunities to study, at atomic resolution, Fabsā?? higher order structures, dynamics and interactions, using solution-state NMR.
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0 Replies | 115 Views
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