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In this new conformation that is predominantly associated with the mutant enzyme, the nicotinamide ring is displaced from its conserved location and three water molecules complete a network of hydrogen bonds between the nicotinamide ring and the protein.
This new conformation was predicted by Rosetta to within 1.2 Å RMSD of the structure determined by X-ray crystallography, representing an unusually accurate structure prediction.
In this new conformation, the PDZ1 domain interacts with one of the protease loops (L3) and becomes locked into the optimal orientation to build up the 12-mer and 24-mer oligomers.
Achieving this new conformation and relieving the intramolecular inhibition of the cargo motif-binding sites are facilitated by large changes occurring to the structure of the whole AP2 core.
In other words, once a stretch of DNA is "trapped" in the transcriptional machinery, the DNA will compact and decompact itself in a way to adapt to this new conformation, and the most efficient way to do so, is by spacing the large loops of DNA evenly.
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In turn, this deprotonation event can stabilize the new conformation of S3.39 relative to D2.50.
Overall, there are two possibilities; either the algorithm reaches the bottom level and finds that the new conformation is within the RMSD cutoff of an existing conformer, in which case it is discarded, or else it is of sufficient diversity to be stored at some level of the tree.
A red shift of 80 nm in the ultraviolet visible absorption maximum suggests a much more extended conjugated π-system for the new conformation.
Those changes could also correspond to a minor modification of tryptophan "embedding" in the new conformation.
Determining Cα RMSD values using the simulation average structure as a reference indicated that although the change in conformation in the side loop varied from the SFTI-1 structure, the new conformation was stable (Figure 2B and C).
It is possible that the crystal forces in the three-dimensional crystal inhibit any significant rearrangement of helices that would normally occur upon illumination, and re-crystallization in the new conformation may be required.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com