Spectroscopic studies on Erythrina lysistemon seed lectin: effect of denaturing agents on lectin quaternary structure.
The equilibrium unfolding of ElysL, a homodimeric legume lectin, was studied using different denaturing agents such as guanidinium chloride (GdnHCl), temperature and pH. Simultaneously, changes in the secondary as well as tertiary structure of lectin were followed by CD spectroscopy examination in both far and near-UV region, respectively. The hydrophobic cluster binding dye, 1-anilino-8-naphthalene sulfonate (ANS), was used to further explore intermediates and to follow the unfolding pathway of lectin. The adenine binding ability of lectin was examined and monitored via absorption spectra and the intrinsic tryptophan fluorescence. Our findings indicate that the ElysL unfolding process occurs via a three state pathway with an intermediate state. We also showed that ElysL binds adenine in a manner that involves a hydrophobic binding pocket that is independent of the carbohydrate binding sites.
Konozy
Emadeldin Hassan E
EH
Structural Biology Unit, Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan 115. Taiwan. ehkonozy@yahoo.com
Dafalla
Maha Babiker
MB
Hsiao
Chwan-Deng
CD
D037121:Plant Lectins / Q000737:chemistry*; D011489:Protein Denaturation / Q000187:drug effects; D020836:Protein Structure, Quaternary / Q000187:drug effects; D012639:Seeds / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
D037121:Plant Lectins; C045629:erythrina lectin