Unraveling the effect of changes in conformation and compactness at the antibody V L -V H interface upon antigen binding
Résumé
We have analyzed conformational changes that occur at the interface between the light (V L) and heavy (V H) chains in antibody variable fragments upon binding to antigens. We wrote and applied the Tiny Probe program that computes the buried atomic contact surface area of three-dimensional structures to evaluate changes in compactness of the V L-V H interface between bound and unbound antibodies. We found three categories of these changes, which correlated with the size of the antigen. Upon binding, medium-sized nonprotein antigens cause an opening of the V L-V H interface (less compact), small antigens or haptens cause a closure of the interface (more compact), whereas large protein antigens have little effect on the compactness of the V L-V H interface. The largest changes in the atomic buried contact surface area at the V L-V H interface occur in residue pairs providing two 'shock absorbers' between the edge b-strands of the V L and V H b-sheets forming the antibody binding site. Importantly, the correlation between the size of antigens and conformational changes indicates that the V L-V H interface in antibodies plays a significant role in the antigen binding process. Furthermore, as the energy involved in such a motion is significant (up to 3 kcal/mol), these results provide a general mechanism for how residues distant from the combining site can significantly alter the affinity of an antibody for its antigen. Thus, mutations introduced at the V L-V H interface can be used to change antibody binding affinity with antigens. Due to the tightly packed V L-V H interface, the introduction of random mutations is not advisable. Rather our analysis suggests that concerted mutations of residues preceding CDRL2 and following CDRH3 or residues preceding CDRH2 and at the end of CDRL3 are most likely to alter or improve antigen binding affinity.