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Reference type: Journal
Authors: Iqbal SS, Lulka MF, Chambers JP, Thompson RG, Valdes JJ
Article Title: Artificial receptors: molecular imprints discern closely related toxins.
Publication date: 2000
Journal: Materials Science & Engineering C-Biomimetic And Supramolecular Systems
Volume: 7
Page numbers: 77-81.
DOI: 10.1016/S0928-4931(99)00090-9

Abstract: We describe silane molecular imprints which can discriminate closely related toxins, supporting the potential usefulness of molecular imprints as artificial receptors, i.e., sensing elements, for detection of biological toxins. Molecular imprints to N-acetylated mu-Conotoxin GIIIB (NacGIIIB) were prepared and binding assays carried out using [C-14]-NacGIIIB. In accordance with a Type I Langmuir adsorption isotherm, saturation of NacGIIIB imprint binding sites occurred at approximately 6.25 mu M and Scatchard analysis revealed a relatively high binding affinity (K-d = 0.37 nM). Binding assays carried out in the presence of a 10-fold molar excess of unlabeled ligand (NacGIIIB) indicated a significant decrease in bound [C-14]-NacGIIIB, i.e., > 90%. In contrast, incubation of [C-14]- NacGIIIB with GIIIA or GIIIB as well as a variety of other Conotoxins demonstrated no significant competition. However, NacGIIIB's closest congener, i.e., NacGIIIA, competed, but required a higher concentration (K-0.5 congruent to 12.5 mu M) to achieve 50% reduction in binding of [C-14]-NacGIIIB than that observed for NacGIIIB (K-0.5 congruent to 6.25 mu M) These results suggest that simple silane imprints can discern subtle differences in tertiary structure of closely related small proteins. (C) 2000 Elsevier Science S.A. All rights reserved


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