Enzymes have been successfully used for detection of analytes providing both recognition and amplification of the binding event with a detectable readout.
Interestingly, for 6His‐tagged G375D, we only detected one binding event with a K D of 0.39 ± 0.10 μM, suggesting that the mutation leads to the loss of the low‐affinity binding site and affects the binding affinity of the high‐affinity binding site.
Genes were defined as targets where a binding event (with FDR < 0.1%) occurred within 5 kb of the transcriptional unit (depending on the proximity of adjacent genes).
Consequently, the composite LysM1 LysM3 binding site provides a single binding event with ultra-high (pM) affinity for chitin binding, the highest chitin-binding affinity described in nature, which is extremely competent to sequester chitin oligosaccharides.
Consistent with this notion, ITC experiments revealed a binding stoichiometry (N) of 0.5, indicating binding of two BRDs to the H4K8acK12ac peptide, whereas only a single binding event with significantly increased affinity was observed for the H4K5acK8ac peptide (Table S8).
Figure 4C shows histograms plotting the number of events where Grb2 docked to another receptor within a 50 second interval after dissociating from a previous binding event, with comparisons in the three spatial environments.
This detailed analysis supports the notion of a global response of the PDZ domain to the binding event, with effects propagated to distal sites, and reveals unexpected roles for the sequences flanking the canonical PDZ domain boundaries.
However, the application of molecular imprinting in the preparation of fluorescent sensors has been hampered by the lack of suitable fluorescent tags, which would respond to the binding event with significant fluorescence intensity changes.
Each overlapping event was tabulated and the numbers of overlaps of each TF's binding event with specific repetitive elements were calculated.
