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These results are consistent with the larger NA activity of the susceptible (WT) strain compared to the H275Y mutant, reported here and by others [25], and its increased NA surface expression [24].
The structure of the H300N mutant reported here (see below) was used in all of the calculations for this variant.
The p Ka values for the substrate-free enzyme were calculated directly from the crystal structure of wild-type StEH1 (PDB entry 2CJP) and the crystal structure of the H300N mutant reported here (PDB entry 4Y9S).
The study of a fresh extract of senescent leaves of the mes16 mutant, reported here, revealed several novel colorless DCCs as the actual major chlorophyll breakdown products in this plant (besides the known FCCs and NCCs[ 21] as minor components), and allowed their structural characterization.
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Contrasting consensus α/β-hydrolases, the bipartite architecture of MHETase parts catalysis from substrate recognition a scenario where we envision the lid domain as a tunable platform to enhance catalytic properties (e.g. alleviating substrate release) or alter substrate specificity (as shown initially for the S416A, R411Q or F424N mutants reported here).
In any case, the decrease in the cellular supply of ATP derived from the ETC is expected to exacerbate at least some of the translation and fidelity defects observed with the mutants reported here.
Furthermore, the hOGT TPR truncation mutants reported here suggest an important function for the TPRs proximal to the active site in the recognition of the large protein substrate TAB1.
Considering the general function of the DnaJ proteins as chaperone proteins [ 1], the reduced amount of Rubisco Activase in the mutants reported here, suggests that the three small chloroplast DnaJ proteins are involved in the folding, unfolding, or assembly processes of this enzyme and thus participate in regulation of Rubisco activity [ 20].
To confirm this finding for a range of TUBA1A mutants reported here and elsewhere, we introduced the mutations I188L, P263T, R264C, L286F, R402H, R402C and S419L into a construct expressing C-terminally FLAG-tagged TUBA1A and transfected these into P19 cells (Fig. 5).
Despite the apparent differences in the effects on spindle length, the overall conclusion that KLP10A plays a role in oocyte spindle length regulation (Zou et al., 2008) is supported by the analysis of klp10A knockdown and loss-of-function mutants reported here and by others (Radford et al., 2012).
Among the structures of the six KaiC mutant proteins reported here, those of the TaT and Tae mutants are the most intriguing because they imply that T426 can be a third phosphorylation site.
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