Functional mapping of conserved residues located at the VL and VH domain interface of a Fab

Journal of Molecular Biology
J ChatellierD Altschuh

Abstract

The interface between the VL and VH domains of antibodies is highly conserved. To investigate the influence of conserved interface residues on Fab function, 13 interface residues were subjected to codon-based combinatorial alanine scanning mutagenesis in Fab 57P, specific for peptide 134 to 151 of the coat protein of tobacco mosaic virus. The 13 single mutants were analysed by Western blot to determine the effect of interface modifications on Fab expression. The kinetic rate constants of peptide-Fab mutant interactions were measured using the biosensor technology. Alanine replacements did not prevent assembly of the mutated Fabs and led to a modification of their binding properties in every case. Twelve of the 13 target residues correspond to homologous positions in the VL and VH domains, which have similar folds. Mutation at homologous positions mostly had different effects on antigen binding affinity. The replacement of bulky side-chains had the most drastic effect on binding. When smaller side-chains were replaced by alanine, the binding properties of Fab mutants differed slightly (by less than a factor of two), but significantly from that of Fab 57P. Modification of some of these residues, which are located 9 to 12 A away f...Continue Reading

Citations

Feb 4, 2010·Protein Engineering, Design & Selection : PEDS·Hiroto IwaiHiroshi Ueda
Jul 2, 2010·Protein Engineering, Design & Selection : PEDS·K R Abhinandan, Andrew C R Martin
Jul 18, 2012·Protein Engineering, Design & Selection : PEDS·Narayan JayaramAndrew C R Martin
Jul 30, 2005·Archives of Biochemistry and Biophysics·Peter J SimonDaniel A Soltis
May 30, 2002·Protein Engineering·Laurence ChoulierDanièle Altschuh
Mar 17, 1999·Journal of Molecular Recognition : JMR·M H Van RegenmortelG Zeder-Lutz
Oct 21, 2003·International Journal of Cancer. Journal International Du Cancer·Michaela A E ArndtSusanna M Rybak
Nov 26, 2002·Journal of Molecular Recognition : JMR·Marc H V Van Regenmortel
Jul 2, 2003·Journal of Molecular Recognition : JMR·Enrique Vargas-Madrazo, Enrique Paz-García
Jul 9, 1999·Journal of Molecular Recognition : JMR·S Muyldermans, M Lauwereys
Jun 27, 2000·Journal of Molecular Recognition : JMR·M B KhalifaT Vernet
Nov 6, 2018·Frontiers in Immunology·Mathieu DondelingerMarylène S Vandevenne
Sep 28, 2017·Scientific Reports·Eva Maria HeroldJohannes Buchner
Jul 12, 2019·Biochimica Et Biophysica Acta. Proteins and Proteomics·Sylvia DietrichGordana Wozniak-Knopp
Oct 16, 2012·Biochemical and Biophysical Research Communications·Danièle AltschuhKarl Andersson
Feb 1, 1997·Molecular Immunology·N Rauffer-BruyreD Altschuh
May 26, 2021·MAbs·Monica L Fernández-QuinteroKlaus R Liedl

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