Dataset Information

Kinetics of Fast Tetramerization of the Huntingtin Exon 1 Protein Probed by Concentration-Dependent On-Resonance R1? Measurements.

ABSTRACT: An approach for the quantitative description of the kinetics of very fast exchange processes (?ex < 50-100 ?s) associated with transient, reversible protein oligomerization, is presented. We show that on-resonance 15N-R1? measurements conducted as a function of protein concentration at several spin-lock radio frequency field strengths are indispensable for unambiguous determination of the rate constants for interconversion between monomeric and higher order oligomeric species. The approach is experimentally demonstrated on the study of fast, reversible tetramerization of the full-length Huntingtin exon 1 protein, httex1, responsible for Huntington's disease. Incorporation of concentration-dependent 15N-R2,eff data, obtained from on-resonance R1? measurements performed at three spin-lock field strengths, into analysis of the kinetic scheme describing reversible tetramerization of httex1 allowed us to uniquely determine the rate constants of interconversion between the various species. This approach serves as a valuable complement to the existing array of NMR techniques for studying early, transient oligomerization events in protein aggregation pathways.

SUBMITTER: Ceccon A

PROVIDER: S-EPMC7377354 | biostudies-literature | 2020 Jul

REPOSITORIES: biostudies-literature

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Kinetics of Fast Tetramerization of the Huntingtin Exon 1 Protein Probed by Concentration-Dependent On-Resonance R1ρ Measurements.

Ceccon Alberto A Tugarinov Vitali V Clore G Marius GM

The journal of physical chemistry letters 20200701 14

An approach for the quantitative description of the kinetics of very fast exchange processes (τex < 50-100 μs) associated with transient, reversible protein oligomerization, is presented. We show that on-resonance 15N-R1ρ measurements conducted as a function of protein concentration at several spin-lock radio frequency field strengths are indispensable for unambiguous determination of the rate constants for interconversion between monomeric and higher orde ...[more]

PMID: 32589032

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Project description:BackgroundFast and accurate T1ρ mapping in myocardium is still a major challenge, particularly in small animal models. The complex sequence design owing to electrocardiogram and respiratory gating leads to quantification errors in in vivo experiments, due to variations of the T1ρ relaxation pathway. In this study, we present an improved quantification method for T1ρ using a newly derived formalism of a T1ρ* relaxation pathway.MethodsThe new signal equation was derived by solving a recursion problem for spin-lock prepared fast gradient echo readouts. Based on Bloch simulations, we compared quantification errors using the common monoexponential model and our corrected model. The method was validated in phantom experiments and tested in vivo for myocardial T1ρ mapping in mice. Here, the impact of the breath dependent spin recovery time Trec on the quantification results was examined in detail.ResultsSimulations indicate that a correction is necessary, since systematically underestimated values are measured under in vivo conditions. In the phantom study, the mean quantification error could be reduced from - 7.4% to - 0.97%. In vivo, a correlation of uncorrected T1ρ with the respiratory cycle was observed. Using the newly derived correction method, this correlation was significantly reduced from r = 0.708 (p < 0.001) to r = 0.204 and the standard deviation of left ventricular T1ρ values in different animals was reduced by at least 39%.ConclusionThe suggested quantification formalism enables fast and precise myocardial T1ρ quantification for small animals during free breathing and can improve the comparability of study results. Our new technique offers a reasonable tool for assessing myocardial diseases, since pathologies that cause a change in heart or breathing rates do not lead to systematic misinterpretations. Besides, the derived signal equation can be used for sequence optimization or for subsequent correction of prior study results.

Project description:BackgroundR1ρ (or spin-lock) imaging is prone to artifacts arising from field inhomogeneities that may impact the R1ρ quantification. Previous research has proposed two types of method to manage the artifacts in continuous-wave constant amplitude spin-lock, one is based on the composite block pulses to compensate for the field imperfections, another category uses adiabatic pulses in the R1ρ pre-pulse to excite and reverse the magnetization (named adiabatic prepared approach). Although both methods have proved their efficiency in alleviating artifacts, we observed that the adiabatic pulse approach could produce much lower R1ρ dispersion in human knee cartilage than the block pulse method (characterized by the R1ρ difference ∆R1ρ =11.4 Hz (from spin-lock field 50 to 500 Hz) for the block pulse method vs. ∆R1ρ =4.5 Hz for the adiabatic pulse approach). Prompted by this observation, the purpose of this study was to investigate the underlying factors that may affect the R1ρ dispersion through numerical simulations based on the two-pool exchanging Bloch-McConnell equations.MethodsThe effects of free water pool size Pa (from 0.80 to 0.95), chemical exchange rate kb (from the bound to free water pool, ranged from 500 to 3,000 Hz), adiabatic pulse duration Tp (from 5.0 to 25 ms), and the chemical shift of the bound pool ppmb (from 1.0 to 5.0 ppm) were examined on the degree of the R1ρ dispersion for the two R1ρ imaging methods.ResultsIn general, the greater the ppmb, kb, Tp, and the smaller Pa, the more significant difference in R1ρ dispersion between the block and adiabatic approaches, with the dispersion curve of the adiabatic method becoming flatter.ConclusionsThe adiabatic prepared approach may compromise the R1ρ dispersion, the effect is determined by the combination of the tissue and radiofrequency (RF) pulse properties. It is suggested that care should be taken when using the adiabatically prepared approach to study R1ρ dispersion.

Dataset Information

Kinetics of Fast Tetramerization of the Huntingtin Exon 1 Protein Probed by Concentration-Dependent On-Resonance R1? Measurements.

Publications

Kinetics of Fast Tetramerization of the Huntingtin Exon 1 Protein Probed by Concentration-Dependent On-Resonance <i>R</i><sub>1ρ</sub> Measurements.

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