Research Papers: Sensing

Fluorescence emission and polarization analyses for evaluating binding of ruthenium metalloglycoclusters to lectins and tetanus toxin C-fragment

[+] Author Affiliations
Tomoko Okada, Norihiko Minoura

Tokyo University of Technology, Graduate School of Bionics, Katakura, Hachioji, Tokyo 194-0982, Japan

J. Biomed. Opt. 16(3), 037001 (March 16, 2011). doi:10.1117/1.3558727
History: Received March 23, 2010; Revised January 04, 2011; Accepted January 27, 2011; Published March 16, 2011; Online March 16, 2011
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We develop a fluorescent ruthenium metalloglycocluster for use as a powerful molecular probe in evaluating the binding between carbohydrates and lectins by fluorescence emission (FE) and fluorescence polarization (FP) analyses. Changes in the FE and FP of these metalloglycoclusters are measured following the addition of lectin [peanut agglutinin (PNA), Ricinus communis agglutinin 120, Concanavalin A (ConA), or wheat germ agglutinin] or tetanus toxin c-fragment (TCF). After the addition of PNA, the FE spectrum of [Ru(bpy-2Gal)3] shows a new emission peak and the FP value of [Ru(bpy-2Gal)3] increases. Similarly, the FE spectrum of [Ru(bpy-2Glc)3] shows a new emission peak and the FP value increases on addition of ConA. Because other combinations of metalloglycoclusters and lectins show little change, specific binding of galactose to PNA and that of glucose to ConA are confirmed by the FE and FP measurements. Resulting dissociation constants (Kd) prove that the metalloglycoclusters with highly clustered carbohydrates show higher affinity for the respective lectins than those with less clustered carbohydrates. Furthermore, specific binding of [Ru(bpy-2Gal)3] to TCF was confirmed by the FP measurement.

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© 2011 Society of Photo-Optical Instrumentation Engineers (SPIE)

Citation

Tomoko Okada and Norihiko Minoura
"Fluorescence emission and polarization analyses for evaluating binding of ruthenium metalloglycoclusters to lectins and tetanus toxin C-fragment", J. Biomed. Opt. 16(3), 037001 (March 16, 2011). ; http://dx.doi.org/10.1117/1.3558727


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