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These results indicate that the upper band of the single mutants denote a sialylated complex N-glycan associated with the N220Q or N229Q Kv3.1 proteins while the lower band is a simple N-glycan.
This mutant, denoted clone B1, contained a non-synonymous mutation (K144R) in gene P3 of the M segment, the locus for the host attachment protein [ 32, 36].
The previously designed T172R/G173Q mutant (denoted as enzyme E172 173) with an improved in vitro half-life of ∼6 h at 37 °C is currently in clinical trial Phase II for cocaine overdose treatment.
The computationally designed new mutant, i.e. the T172R/G173Q/L196C/I301C mutant (denoted as enzyme E196 301 for convenience), may have a pair of cross-subunit disulfide bonds: one between C196 of subunit a (C196a) and C301 of subunit b (C301b), and the other between C301 of subunit a (C301a) and C196 of subunit b (C196b).
The T172R/G173Q mutant (denoted as enzyme E172 173 here for convenience) was designed through introducing favorable noncovalent forces including a hydrogen bond between domains I and II of the protein.
Plus denotes a mutant allele, whilst minus denotes wild-type.
In these matrices, G u, v quantifies a phenotype of double mutant u Δ v Δ and S u denotes a phenotype of single mutant u Δ.
Here * denotes a mutation.
An asterisk denotes a statistically significant difference between wild type and gs1 2 mutants (*, P < 0.05 by Student's t-test).
* denotes a highly recommended show.
* denotes a highly recommended concert.
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