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Strategies for acquisition of resonance assignment spectra of highly dynamic membrane proteins: a GPCR case study
Jul 27, 2023 - 10:14 PM - by nmrlearner
nmrlearner's Avatar Strategies for acquisition of resonance assignment spectra of highly dynamic membrane proteins: a GPCR case study

Abstract

In protein nuclear magnetic resonance (NMR), chemical shift assignment provides a wealth of information. However, acquisition of high-quality solid-state NMR spectra depends on protein-specific dynamics. For membrane proteins, bilayer heterogeneity further complicates this observation. Since the efficiency of cross-polarization transfer is strongly entwined with protein dynamics, optimal temperatures for spectral sensitivity and resolution will depend not only on inherent protein dynamics, but temperature-dependent phase properties of the bilayer environment. We acquired 1-, 2-, and 3D homo- and heteronuclear experiments of the chemokine receptor CCR3 in a 7:3 phosphatidylcholine:cholesterol lipid environment. 1D direct polarization, cross polarization (CP), and T2â?? experiments indicate sample temperatures belowâ??â??â??25Â*°C facilitate higher CP enhancement and longer-lived transverse relaxation times. T1rho experiments indicate intermediate timescales are minimized below a sample temperature ofâ??â??â??20Â*°C. 2D DCP NCA experiments indicated optimal CP efficiency and resolution at a sample temperature ofâ??â??â??30Â*°C, corroborated by linewidth analysis in 3D NCACX atâ??â??â??30Â*°C compared toâ??â??â??5Â*°C. This optimal temperature is concluded to be directly related the lipid phase transition, measured to be betweenâ??â??â??20 and 15Â*°C based on rINEPT signal of all-trans and trans-gauche lipid acyl conformations. Our results have critical implications in acquisition of SSNMR membrane protein assignment spectra, as we hypothesize that different lipid compositions with... [Read More]
0 Replies | 358 Views
A direct nuclear magnetic resonance method to investigate lysine acetylation of intrinsically disordered proteins - Frontiers
Jul 27, 2023 - 10:14 PM - by nmrlearner
nmrlearner's Avatar A direct nuclear magnetic resonance method to investigate lysine acetylation of intrinsically disordered proteins Frontiers
A direct nuclear magnetic resonance method to investigate lysine acetylation of intrinsically disordered proteins - Frontiers
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0 Replies | 169 Views
Researchers investigate how the cellular environment affects protein conformational dynamics - Phys.org
Jul 27, 2023 - 10:14 PM - by nmrlearner
nmrlearner's Avatar Researchers investigate how the cellular environment affects protein conformational dynamics - Phys.org

Researchers investigate how the cellular environment affects protein conformational dynamics Phys.org Read here
0 Replies | 222 Views
Chinese Academy Explores Protein Dynamics in Live Cells - Mirage News
Jul 27, 2023 - 9:43 AM - by nmrlearner
nmrlearner's Avatar Chinese Academy Explores Protein Dynamics in Live Cells - Mirage News

Chinese Academy Explores Protein Dynamics in Live Cells Mirage News Read here
0 Replies | 170 Views
An expandable, modular de novo protein platform for precision ... - pnas.org
Jul 26, 2023 - 5:24 PM - by nmrlearner
nmrlearner's Avatar An expandable, modular de novo protein platform for precision ... - pnas.org

An expandable, modular de novo protein platform for precision ... pnas.org Read here
0 Replies | 155 Views
Global Protein Crystallization And Crystallography Market Size and ... - Glasgow West End Today
Jul 25, 2023 - 1:43 PM - by nmrlearner
nmrlearner's Avatar Global Protein Crystallization And Crystallography Market Size and ... - Glasgow West End Today

Global Protein Crystallization And Crystallography Market Size and ... Glasgow West End Today Read here
0 Replies | 187 Views
[NMR paper] NMR structure verifies the eponymous zinc finger domain of transcription factor ZNF750
Jul 25, 2023 - 1:43 PM - by nmrlearner
nmrlearner's Avatar NMR structure verifies the eponymous zinc finger domain of transcription factor ZNF750

ZNF750 is a nuclear transcription factor that activates skin differentiation and has tumor suppressor roles in several cancers. Unusually, ZNF750 has only a single zinc-finger (ZNF) domain, Z*, with an amino acid sequence that differs markedly from the CCHH family consensus. Because of its sequence differences Z* is classified as degenerate, presumed to have lost the ability to bind the zinc ion required for folding. AlphaFold predicts an irregular structure for Z* with low confidence. Low...

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0 Replies | 396 Views
[NMR paper] Imaging Saturation Transfer Difference (STD) NMR: Affinity and Specificity of Protein-Ligand Interactions from a Single NMR Sample
Jul 25, 2023 - 1:43 PM - by nmrlearner
nmrlearner's Avatar Imaging Saturation Transfer Difference (STD) NMR: Affinity and Specificity of Protein-Ligand Interactions from a Single NMR Sample

We have combined saturation transfer difference NMR (STD NMR) with chemical shift imaging (CSI) and controlled concentration gradients of small molecule ligands to develop imaging STD NMR, a new tool for the assessment of protein-ligand interactions. Our methodology allows the determination of protein-ligand dissociation constants (K(D)) and assessment of the binding specificity in a single NMR tube, avoiding time-consuming titrations. We demonstrate the formation of suitable and reproducible...

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0 Replies | 214 Views
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