In the present study, we investigated the spectroscopic properties and photoreaction processes of mutants of the KR2 NDQ motif.
The process of mutant protein folding is particularly sensitive to such insults.
Hrd1 (21, 34) and gp78 (35) have also been well-characterized in the degradation process of mutant alpha-1-antitrypsin and many other misfolded proteins (36, 37).
Typical chemical chaperones are small molecules (e.g. glycerol, dimethyl sulfoxide, 4-phenylbutyrate, trimethylamine) that stabilise the folding process of mutant proteins and/or the assembly of multimeric complexes by increasing hydration of the folding mutant (Ref. 90).
Although mutated proteins sometimes induce ER stress before being degraded (Vidal et al, 2011), the expression level of ER-stress-responsive molecules was comparable between the cells expressing ZIP13WT and the pathogenic mutants (Supplementary Fig S11), indicating that ER stress might not significantly participate in the pathogenic process of mutant ZIP13 proteins.
Immunostaining for YFP and Phalloidin to visualize the actin cytoskeleton in similar cultures (Fig. 5 C D′), revealed extensive splitting of the leading processes of double mutant cells after 2 DIV, a phenotype that is not frequently observed in control or single Rac1 conditional mutants (Fig. 5 C′,D′ and Fig. 6 A).
This technical repeat therefore only quantifies variation associated with sample preparation and sequencing, whereas the biological replicates also quantify variation associated with the processes of codon-mutant library creation, virus generation, and virus passage.
Directed evolution of enzymes consists of an iterative process of creating mutant libraries and choosing desired phenotypes through screening or selection until the enzymatic activity reaches a desired goal.
To further validate our model, we also investigated the labeling process of two mutant strains, GDH knockout (ΔGDH) and GOGAT knockout (ΔGOGAT).
