PMID: 11932494Apr 5, 2002Paper

Mapping of a hapten-binding site: molecular modeling and site-directed mutagenesis study of an anti-atrazine antibody

Protein Engineering
Wien KusharyotoRolf D Schmid

Abstract

A three-dimensional model of the variable domain of the atrazine-specific Fab fragment K411B was constructed by molecular modeling using known structures of highly homologous immunoglobulins as templates. Molecular dynamic simulations and cross-reactivity data were used to predict residues responsible for the binding of the hapten 4-chloro-6-(isopropylamino)-1,3,5-triazine-2-(6-aminohexanecarboxylic acid) (iPr/Cl/C6) instead of atrazine. Specific binding pockets could be defined for the chlorine, the isopropylamino group and the C6-spacer of the hapten. The influence of various amino acids on hapten binding was investigated by site-directed mutagenesis, and the effect of these mutations was analyzed by capture ELISA using the hapten iPr/Cl/C6 and 4-amino-6-chloro-1,3,5-triazine-2-(6-aminohexanecarboxylic acid) (H/Cl/C6). GlyH100a seems to be important in determining the conformation of the heavy-chain complementarity determining region H3; replacing it with any other residue prevented the binding of the hapten. Altering residues responsible for the binding of the chlorine atom (TrpH33, GluH50 and TyrL96) decreased the affinity significantly. Hapten-spacer recognition can be attributed to the interaction with PheL32; replacing P...Continue Reading

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Citations

Mar 12, 2003·Prostaglandins, Leukotrienes, and Essential Fatty Acids·Lilian Rumi TsurutaMichinao Mizugaki
Apr 30, 2008·BMC Bioinformatics·Maria Pamela C DavidVincent Ricardo M Daria
Feb 23, 2011·International Journal of Molecular Sciences·Jian-Xin ZhaoWei Chen
Mar 27, 2007·Biotechnology Advances·Claudia SheedyJ Christopher Hall
Sep 15, 2012·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Xiaobo LiuHiroshi Ueda
Jan 21, 2004·Journal of Molecular Recognition : JMR·Rebecca L Rich, David G Myszka
Jul 30, 2020·Protein Engineering, Design & Selection : PEDS·Daniel A McPartlinRichard J O'Kennedy
Apr 18, 2019·International Journal of Molecular Sciences·Chia Chiu LimTheam Soon Lim

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