Determination of affinities of oligopeptides binding with p65 and identification of their inhibiting effect on DNA binding activity of NF-?B / 第三军医大学学报

ABSTRACT

Objective To determine the affinities of five oligopeptides specifically binding with DNA binding domain of NF-?B p65 subunit and identify their inhibiting effect on DNA binding activity of NF-?B. Methods By using biosensor the affinities were measured by means of kinetic analysis, and the inhibiting effects were determined by competitive ELISA. Results The results of biosensor showed that all of five oligopeptides really possessed the capability of specific interacting with NF-?B p65 subunit. The affinity constants of these oligopeptides were 2.67?10~ -7 mol/L, 9.02?10~ -6 mol/L, 1.07?10~ -6 mol/L, 8.03?10~ -6 mol/L, 9.83?10~ -7 mol/L respectively. The results of competitive ELISA indicated that five oligopeptides could inhibit NF-?B from binding with ?B motif, and their inhibiting effect depended on their concentration. Conclusion Five oligopeptides that were screened by yeast two-hybrid system method can really interact with p65, and possess the inhibiting effect on DNA binding activity of NF-?B. So it will be possible that these oligopeptides are regarded as model to design and develop novel anti-inflammatory peptide drug targeting NF-?B.