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A Simple Fluorescence-based Reporter Assay to Identify Cellular Components Required for Ricin Toxin A Chain (RTA) Trafficking in Yeast
Published on: December 15, 2017
A novel sugar-probe biosensor for the deadly plant proteinous toxin, ricin
Hirotaka Uzawa1, Koji Ohga, Yukiko Shinozaki
1Research Center of Advanced Bionics, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba 305-8565, Japan.
Abstract:
Because of the illegal use of highly toxic ricin from the castor-oil plant, Ricinus communis, in bioterrorism and suspected white powder cases, anti-terrorism measures for the toxin are urgently required. Here we demonstrate a facile and sensitive detection method using synthetic analogues of beta-lactosyl- and beta-d-galactosyl ceramides as the ligands based on the fact that ricin binds cell-surface oligosaccharides. Sugar-probes having lipoic acids as anchor functions were synthesized via either a chemical or chemoenzymatic way and were immobilized on the sensor chips by a self-assembled monolayer technique. Surface plasmon resonance (SPR) analysis using these carbohydrate probes allowed us to detect the toxin in a highly sensitive and facile manner (10 pg/mL, 5 min), being the best benchmark as a method for detecting the toxin. In addition, a visual monitoring method was developed, in which sugar-coated Au nanoparticles were utilized for discriminating ricin from other proteins in a facile manner, taking 10-30 min for judgment.

