(C ) alpha backbone mutations.
A previous study has shown that a backbone L184 mutation causes partial loss of function and converts the partial agonist mCPBG into an antagonist.
A variant of elotuzumab was generated with an IgG2 backbone and mutations in the Fc region (Elo IgG2M3) as previously described [ 29].
Indeed, crenezumab, which uses an IgG4 backbone with mutations that reduce affinity for FcR [ 32], appeared in the initial phase I study to avoid amyloid-related imaging abnormalities (ARIAs) even at higher doses than have been tolerated for other anti-Aβ mAbs.
The response of recombinant 3′-UTR motifs to miR-181b was normalised with respect to a mutant miR-181b control (miR-181b_mut, designed from the miR-181b backbone with mutations introduced at positions 3, 5, and 7; see Additional file 1: Table S1 for oligonucleotide sequences).
Additionally, we calculated the change in root-mean-square fluctuation of all Cα atoms as a measure of altered backbone flexibility following mutation according to ΔRMSF = RMSF (mutant) — RMSF (wild-type).
The limited number of mutations observed in the genome backbone are consistent with mutation rates in bacteria previously calculated using data from both experimental (e.g., [ 2, 3]) and natural populations [ 38].
Mass spectroscopy measurements also showed that the ΔF508 mutation increased backbone dynamics at residues 509−511.
DN-Akt (Addgene, plasmid 9031) was built by generating a vector backbone (pcDNA3.0) with a mutation that deprived Akt of its phosphorylation ability.
However, such an approach fails for HDAC8, as the potassium is mostly coordinated by the protein backbone, and also because mutation of side chains that do participate in the coordination of K+ (S199 and D176, see Figure 1 C)[ 15] resulted in insufficient purified protein for further NMR characterisation.
Using the assumption that the hGBE1-apo crystal structure represents an active enzyme conformation, the design of an hGBE1 p.Y329S-stabilizing peptide was performed using a rigid backbone modelling of the mutation, in order to retain maximum similarity to the active enzyme.
