Artículos de revistas
An unusual cysteine VL87 affects the antibody fragment conformations without interfering with the disulfide bond formation
Fecha
2017-10Registro en:
Attallah, Carolina Veronica; Aguilar, Maria Fernanda; Garay, Alberto Sergio; Herrera, Fernando Enrique; Oggero, Marcos; et al.; An unusual cysteine VL87 affects the antibody fragment conformations without interfering with the disulfide bond formation; Pergamon-Elsevier Science Ltd; Molecular Immunology; 90; 10-2017; 143-149
0161-5890
CONICET Digital
CONICET
Autor
Attallah, Carolina Veronica
Aguilar, Maria Fernanda
Garay, Alberto Sergio
Herrera, Fernando Enrique
Oggero, Marcos
Rodrigues, Daniel Enrique
Resumen
The Cys residues are almost perfectly conserved in all antibodies. They contribute significantly to the antibody fragment stability. The relevance of two natural contiguous Cys residues of an anti-recombinant human-follicle stimulation hormone (rhFSH) in a format of single-chain variable fragment (scFv) was studied. This scFv contains 5 Cys residues: VH22 and VH92 in the variable heavy chain (VH) and VL23, VL87 and VL88 in the variable light chain (VL). The influence of two unusual contiguous Cys at positions VL87 and VL88 was studied by considering the wild type fragment and mutant variants: VL-C88S, VL-C87S, VL-C87Y. The analysis was carried out using antigen-binding ability measurement by indirect specific ELISA and a detailed molecular modeling that comprises homology methods, long molecular dynamics simulations and docking. We found that VL-C87 affected the antibody fragment stability without interfering with the disulfide bond formation. The effect of mutating the VL-C87 by a usual residue at this position like Tyr caused distant structural changes at the VH region that confers a higher mobility to the VH-CDR2 and VH-CDR3 loops improving the scFv binding to the antigen.