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Exact(5)
The complete assignment of the backbone amide resonances of both proteins [11], [12] allows for a residue-specific analysis of the proton exchange rates (Fig. S7, S8 and Table S1).
The assigned backbone amide resonances of the N,H-HSQC spectrum are shown in Figure 1.
Although we did not see a chemical shift perturbation of the backbone amide resonances of V351, close contact does not necessarily produce shift changes.
Resonance assignments of backbone and side-chain atoms were nearly complete (>97%) with only the H/N backbone amide resonances of V55 unobservable.
Addition of a H3K36me3 peptide (residues 28 to 41) resulted in clear chemical shift changes for the backbone amide resonances of residues around the aromatic cage and strand β4 of the PWWP domain.
Similar(55)
Backbone and Cβ assignments were obtained for all resonances, except for the amide resonances of Ser, which is presumably broadened beyond detection due to rapid hydrogen exchange as observed for the homologous position in FKBP12 [ 54] and FKBP52 [ 43].
This rigidification of the Tyr side-chain mobility is consistent with the reduced conformational dynamics of the Tyr backbone that is indicated by the marked narrowing of the amide resonance of this residue in the H87V NMR spectra.
Most of the backbone amide resonances are visible in the reference spectra of both proteins; however, while 90 out of the 257 amide resonances of scPCNA are still clearly observed after ∼1 h in D2O, only 15 of the 260 cross-peaks of hPCNA could be seen under similar conditions.
Assignment of the backbone amide group resonances of NT* was obtained on the basis of NTwt assignments at pH 7.2 (ref. 32) by analysing a 3D 15N-resolved NOESY-HSQC spectrum acquired with a 60 ms mixing time.
Exchange between the conformers within the monomer ensemble on the microsecond to millisecond timescale renders the majority of backbone amide resonances broadened beyond detection.
In addition, loop 166 172, which is lack of backbone amide resonances in hPrPC and mPrPC, is well-defined in both hPrPC (S170N) and mPrPC (V166A) mutants [16], [41].
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