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To investigate the role of sterol binding, we made a non-sterol-binding mutant by mutating a conserved glycine residue in the C-terminal helix predicted to interact with the omega-1 loop.
While our data do not allow us to explain the role that the other group of Met311 may play in Na+ binding, we made a relevant comment in the revised manuscript.
When we further defined the Tip110 regions that were responsible for the snRNA U6 binding, we made a series of deletion mutants which including the C-terminal (1 350aa), deletion of NSL (∆600 670aa), deletion of two RRM (∆740 850aa).
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Since we had already shown that a D2447N mutation increased binding significantly (144%), we made a triple mutant (D2385K/D2386N/D2447N) but this bound no better than the D2447N mutant alone.
To help us better interpret the Regulus results, and to further explore the binding site content of the CRMs, we made a graphic map of the identified motifs along the sequences for several pairs of CRMs (from this study as well as from the literature; fig. 8 and supplementary fig. S4 a e, Supplementary Material online).
Valine 2376 in the loop contacts W2389 in the centre of the actin-binding site, and we therefore made a V2376D mutant to study the effects on binding of destabilising this loop.
While the Big Innovation Centre's report, the first of a series, rejected pay ratios and binding votes, it made a series of recommendations to tackle excess pay.
Although not contractually binding, the DHBs have made a commitment to split night shifts and to reduce the roster to 10 days.
And we made it legally binding," he added.
A binding contract makes an eviction easier.
Hobbes thinks that we incur binding obligations when we make pledges under the appropriate conditions.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com