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A generated variant containing a Ser residue at this position, however, resulted in poor ability to express soluble and enzymatically active protein.
We developed an ion-pair reversed-phase HPLC method for the separation of an intact monoclonal antibody variant containing a single isoaspartic acid residue from its native counterpart.
These screens identified a glycine to serine mutation of L30e, which specifically restores activity to an RNA variant containing a U that replaces a helix-capping G. Similarly, an asparagine to alanine mutation was found to suppress a substitution at a position where the L30e RNA nucleotide extends out into the protein pocket.
Electrophysiology and structural studies were performed on an annexin V variant containing a mutation of glutamic acid-95 to serine in the center of the pore region.
In contrast, for the growth curve of the OmpA-177 variant containing a FLAG insertion in loop 3, no lag phase is observed after induction with IPTG.
Interestingly, a recent paper shows that, in keeping with this hypothesis, synthetic polyQ peptides of non-pathological length (e.g., KKQ32KK) are much more prone to aggregation than a huntingtin exon 1 variant containing a Q53 stretch [36].
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The Jmjd6-(AxA) variant containing an inactivated JmjC-domain was obtained by replacing the JmjC domain and C-terminal part of Jmjd6 using the product of primer pair 2. The forward primer contains two mutations to inactivate the HxD motif of Jmjd6 (HID changed to AIA in the predicted catalytical triad).
Sequence analysis showed that this variant contained a single point mutation (Leu71Ile).
The variant contained a 131-bp insertion, which was derived from the middle part of intron 1 (9,064-bp downstream of exon 1), between exons 1 and 2 (Figure 2c).
The asymmetric unit for each variant contained a single monomer.
TNFR2 lies within the Idd9 interval, and the diabetes-associated variant contains a mutation close to the TRAF2 binding site.
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