Accordingly, the introduction of a hydroxyl group at the para-position of the benzene ring in vesamicol markedly reduced the affinity for sigma-1 receptor.
ITC analysis indicated that the presence of each of these mutations markedly reduced the affinity of these proteins for PET-16 as well as PES.
Both these estrogen metabolites bind to the estrogen receptor, but with a markedly reduced binding affinity as compared with estradiol [ 37- 39].
As is shown in Figure 1(c), overexpression of DDAH2 in VSMCs markedly reduced the affinity precipitation of RhoA to a normal level by comparing to VSMCs infected with the blank vector pAV-CMV-GFP (P < 0.05).
Analogues that had no effect in the functional assay showed markedly reduced binding affinities.
W430A had strongly reduced affinity for tetramethylammonium.
In contrast, mutant R22E with markedly reduced GAG-binding affinity, had a minimal effect on HUVEC proliferation.
In contrast, mutant R22E, a partial CXCR3 agonist, but with markedly reduced GAG binding affinity, had a minimal effect on HUVEC proliferation even at the highest concentration tested.
Indeed, fluorescence spectroscopy and ITC assays showed that the alanine mutations R2A in the H3 peptide and D312A in AIRE PHD1 markedly reduced the binding affinity (Table 1; Fig 4C) without affecting the protein fold (supplementary Fig S3 online).
These data suggest that peptide residues in the vicinity of the glycosylation site (±5 residues) can enhance glycopeptide affinity, and also that loss of the glycan from the peptide, by enzymatic cleavage, would markedly reduce the affinity of the products.
