jz0c01636_si_001.pdf (1.48 MB)
Kinetics of Fast Tetramerization of the Huntingtin Exon 1 Protein Probed by Concentration-Dependent On-Resonance R1ρ Measurements
journal contribution
posted on 2020-07-02, 00:18 authored by Alberto Ceccon, Vitali Tugarinov, G. Marius CloreAn
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 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 R1ρ 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.
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Concentration-Dependent On-Resonanc...on-resonance R 1ρ measurementsorder oligomeric specieson-resonance 15 NoligomerizationHuntingtin Exon 1 Protein Probedspin-lock radio frequency field str...protein aggregation pathwaysR 1ρ measurementsrate constantsapproachNMRHuntingtin exon 1 proteintetramerizationinterconversionhttspin-lock field strengthsconcentration-dependent 15 N
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