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Under a multiple turnover condition, both performed well at 37°C, showing the kcat of 1 and 0.26 min−1, respectively.
Radioactive assays were carried out at 30°C under a single turnover condition with 5 nM substrate (see Text S1 for substrate preparation) and 1 µM DNAzyme in 70 mM HEPES (pH 8.0) containing 40 mM MgCl2, 0.001% Tween-20.
Notably, low Mg levels and low Mg dietary intake was associated with osteoporosis, a classic high bone turnover condition [ 11].
The inhibin-poor environment of postmenopausal women (as in the AZURE, Z-FAST and ZO-FAST trials) or ovarian-suppressed women (as in the ABCSG12 trial) can therefore support the high bone turnover condition in which ZA is most effective (as discussed above in the context of estrogen deprivation).
During these competition experiments, we noted that overall RIDD cleavage (either with INSULIN or BLOS1 RNA) was rather less effective than HAC1 or XBP1 RNA cleavage, suggesting that 3 μM yIre1 might not truly represent a single turnover condition for RIDD cleavage.
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Confocal microscopy analysis revealed that the thickness of biofilms under turnover conditions and non-turnover conditions was approximately 35 and 3.5 μm, respectively.
G. sulfurreducens biofilms were grown in minimal medium with acetate as electron donor (turnover conditions), followed by 24 h incubation in electron donor-depleted medium (non-turnover conditions).
Probing the enzyme under turnover conditions showed mechanistic insights into proton translocation coupled to electron transfer.
Studies using static samples will be discussed first, before the more technologically challenging in situ studies under turnover conditions.
Further, the protein binds Zn II) and Co II) in the expected manner and shows ferroxidase activity under single turnover conditions.
In this study, we made quantitative measurements of the DNA glycosylase activity of Pa-AGOG wild type and some engineered variants under single turnover conditions.
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