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Deuterium exchange experiments of DARPins with two and three internal repeats between N- and C-terminal capping repeats (NI2C, NI3C) and NI3C_Mut5, where the C-cap had been reengineered, indicate that the stability of the full-consensus ankyrin repeat proteins is strongly dependent on the coupling between repeats, as the stabilized cap decreases the exchange rate throughout the whole protein.
The time scale of removal during in-situ imaging at pH 1 is consistent with the pH-jump experiments of Samson et al.[4] for dissolution of kinetically labile Fe from the hematite surface during approach to steady-state dissolution, and is roughly consistent with the time scale of isotopic exchange experiments of Rea et al.[7] for kinetically labile surface Fe on ferrihydrite.
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For that, we clarified the H-desorption mechanism by the isotopic exchange experiments, on the basis of which we designed a new Li Mg N H composite system with the reaction 8LiH + 3Mg(NH2)2 ↔ 4Li2NH + Mg3N2 + 8H2.
We performed a peptide exchange experiment for each non-redundant set of interaction pairs with 10 or more representative 3D structures, and where at least one peptide was sufficiently long for studies of contextual effects (≥10 residues).
For hydrogen/deuterium (H/H) solvent exchange experiments, 500 μL of a 1 mM N-labeled-IsdBN1 solution previously lyophilized was resuspended in 500 μL of 100% D2O and 2D H N correlation HSQC spectra collected at subsequent times.
We have also performed partial cation exchange experiments, using substoichiometric amounts of Cu+ ions, in order to obtain information about the initial stages of the CE process (see "Methods" section and Supplementary Notes 2).
By combining the knowledge gathered from the internal exon exchange experiments and the modification of the endogenous transcript, we are now planning a new experiment in which we target an internal exon of endogenous mutated nebulin transcripts in vivo using a zebrafish model for nemaline myopathy.
For the fast-solution exchange experiments, the time course of Na+/K+ exchange, pH gating and QA blocking and unblocking was derived from a monoexponential fit.
The kinetic observations taken together with the metal exchange experiments show the advantage of the shuttle mechanism over the displacement mechanism.
The corresponding length scales may be probed directly by two-dimensional T2– T2 relaxation exchange experiments where the appearance of cross-peaks demonstrates that during the measurement time (texch) molecules have moved from a liquid phase to a vapor phase.
The surprising destabilization of β-sheet residues upon nucleotide binding, as seen in hydrogen/deuterium exchange experiments, shows that the number of closest neighbors does not fully explain residue flexibility.
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