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The decrease in recovery at pH values lower than 5.0 may be due to the competition of proton with cations in binding to donor atoms.
Two positively-charged CAD residues, Arg243 and Lys245, are responsible for binding to donor dsDNA [7], [8], [9], [10].
Hence we tested BMT recipients for the presence of antidonor antibodies early (1 and 2 weeks) and late (>3 months) after BMT through flow cytometric assessment of recipient serum binding to donor cells.
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Thus these mutations do not seem to affect the binding to the donor substrate.
This figure corresponds to situations where the reactant (product) electronic charge distribution strongly favors proton binding to its donor (acceptor).
The Km,app-tyramine value for the S182A/S186A double mutant (190 μM) is significantly higher than the wild-type value (12 μM), suggesting that Ser-182 and Ser-186 function synergistically in maintaining the proper active site architecture for optimal binding to the amino donor substrate.
Following the recognition of and binding to the 5´ss (upstream donor site) by the U1 snRNP, the U2 is recruited to the 3´ss (downstream acceptor) and recognizes the BPS, resulting in the formation of the pre-spliceosome.
Control experiments with acceptor-free PEG-coated surface were performed in order to ensure that non-specific binding of donor vesicles was rare; furthermore, we had previously performed a control experiment with a soluble synaptobrevin fragment to disrupt SNARE complex formation which resulted in loss of donor vesicle binding to surface-tethered acceptor vesicles (Kyoung et al., 2011).
The inefficient recovery of cyanide was likely caused upon addition to whole blood by its rapid transformation to volatile HCN gas at pH values below its p Ka of 9.2, enzyme-catalyzed conversion to SCN– in the presence of a sulfur donor, and binding to blood components, including hemoglobin (Hb), methemoglobin (metHb), and albumin.
The DNaseI sensitivity of the remodeled filament indicates that the ssDNA is more accessible, and it is therefore possible that the remodeling induced by Rad51 paralogs exposes the ssDNA to facilitate homology probing after filament binding to and disruption of the donor duplex.
In fact, the only documented 'cure' of HIV infection remains in the German AIDS patient who underwent BM transplantation (BMT) from a CCR5 Δ32 homozygous donor (with decreased binding to R5-tropic HIV strains) as treatment for acute leukemia.
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