Thermodynamic and molecular dynamic insights into how fusion influences peptide-tag recognition of an antibody.

ABSTRACT: To understand the effect of protein fusion on the recognition of a peptide-tag by an antibody, we fused a CCR5-derived peptide-tag (pep1) to GFP and investigated its recognition by an anti-pep1 antibody, 4B08. First, to characterize the thermodynamic properties associated with the pep1-4B08 binding, isothermal titration calorimetry experiments were conducted. It was found that pep1 fused to the C-terminus of GFP (GFP-CT) enhanced the enthalpic gain by 2.1 kcal mol^-1 and the entropic loss only by 0.9 kcal mol^-1, resulting in an 8-fold increase in the binding affinity compared to the unfused pep1. On the other hand, pep1 fused to the N-terminus of GFP (GFP-NT) enhanced the enthalpic gain by 3.0 kcal mol^-1 and the entropic loss by 3.2 kcal mol^-1, le

SUBMITTER: Miyanabe K 

PROVIDER: S-EPMC11018781 | biostudies-literature | 2024 Apr

REPOSITORIES: biostudies-literature