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Bacterial expression, purification and biophysical characterization of wheat germ agglutinin and its four hevein‐like domains
Author(s) -
Leyva Eduardo,
MedranoCerano Jorge L.,
CanoSánchez Patricia,
LópezGonzález Itzel,
GómezVelasco Homero,
RíoPortilla Federico,
GarcíaHernández Enrique
Publication year - 2019
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.23242
Subject(s) - wheat germ agglutinin , chemistry , recombinant dna , lectin , folding (dsp implementation) , biochemistry , agglutinin , binding domain , fusion protein , biophysics , microbiology and biotechnology , binding site , biology , gene , electrical engineering , engineering
Wheat germ agglutinin (WGA), a chitin binding lectin, has attracted increasing interest because of its unique characteristics such as conformational stability, binding specificity and transcytosis capacity. To pave the way for the study of the molecular basis of WGA's structural stability and binding capacity, as well as to facilitate its use in biomedical and biotechnological developments, we produced recombinant WGA and its 4 isolated hevein‐like domains in a bacterial system. All the proteins were expressed as fusion constructs linked to a thioredoxin domain, which was enzymatically or chemically released. The structural and ligand‐binding properties of recombinant WGA were similar to the wild lectin. The 4 isolated domains folded and were ligand‐binding competent, indicating that each domain constitutes an independent folding unity. The biophysical characterization of the recombinant domains sheds new light on the intricate folding and binding behavior of this emblematic lectin.