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The location of the C1 atom of Gal p near the N5-FAD atom is consistent with mechanistic studies showing that the N5-FAD atom functions as a nucleophile.
In this case it functions as a nucleophile.
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Consider how water, because of its lone electron pairs, can function as a nucleophile.
Similarly, Cys can function as a powerful nucleophile in the active sites of enzymes operating under nucleophilic catalysis.
Usually, bases can function in addition to their being bases they can function as nucleophiles as well.
Furthermore, analogous to BVP, β-phosphate hydrolysis by PIR1 proceeds efficiently in the absence of γ-phosphate, arguing against a model in which the Cys−γ-Pi intermediate functions as the nucleophile that attacks the β-phosphorus of an RNA substrate.
In several GH13 enzymes, this constellation of three acidic residues functions as the nucleophile (Asp357, hGBE1 numbering hereinafter), proton donor (Glu412) and transition state stabilizer (Asp481) in the active site.
Indeed, it is not clear that Glu63* functions as nucleophile only, or that a competing pathway exists in which it functions as general base (Scheme 2).
Both of the two catalytic His residues are critical in the reaction course, where one acts as a nucleophile, while the other functions as a general acid/base.
DNA glycosylases that use an amino group of a lysine side chain as a nucleophile for base cleavage form a covalent Schiff-base intermediate with the substrate.
However, the side reactions in the presence of TCEP are usually slower because it has no thiol group that functions as nucleophile competing with trans-splicing reaction.
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