Elbow flexibility and ligand-induced domain rearrangements in antibody Fab NC6.8: large effects of a small hapten

Biophys J. 2000 Aug;79(2):614-28. doi: 10.1016/S0006-3495(00)76320-X.

Abstract

Four 700-ps molecular dynamics simulations were carried out to analyze the structural dynamics of the antigen-binding antibody fragment NC6.8, which is known to exhibit large structural changes upon complexation. The first simulation was started from the x-ray structure of the uncomplexed Fab and produced trajectory averages that closely match the crystallographic results. It allowed assessment of the flexibility of the Fab, revealing an elbow motion of the variable domains with respect to the constant domains. The second simulation was started from the uncomplexed x-ray structure after insertion of the ligand into the binding site. This perturbation resulted in a significantly altered trajectory, with quaternary structural changes corresponding in many aspects to the experimental differences between complexed and uncomplexed state. The observed trend toward a smaller elbow angle and a higher flexion of the H-chain could also be seen in the third simulation, which was started from the x-ray structure of the complex. The changes were revealed to be a clear consequence of the complexation with the ligand because in the fourth simulation (started from the experimental complex structure after removal of the hapten) the Fab remained close to its initial structure. Analyses of the quaternary structure and the binding site of Fab NC6.8 are presented for all four simulations, and possible interpretations are discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antigens
  • Binding Sites, Antibody
  • Computer Simulation
  • Crystallography, X-Ray
  • Haptens*
  • Immunoglobulin Fab Fragments / chemistry*
  • Immunoglobulin Heavy Chains / chemistry
  • Ligands
  • Models, Molecular
  • Protein Conformation
  • Stress, Mechanical
  • Thermodynamics

Substances

  • Antigens
  • Haptens
  • Immunoglobulin Fab Fragments
  • Immunoglobulin Heavy Chains
  • Ligands