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A Cost-Effective Method for the Optical Transduction of Chemical Reactions. Application to Hyaluronidase Inhibitor Screening with Polyarginine-Counteranion Complexes in Lipid Bilayers

Authors
Miyatake, Tomohiro
Published in Journal of the American Chemical Society. 2006, vol. 128, no. 38, p. 12420-12421
Abstract We report the systematic elaboration of a cost-effective, interference-minimized assay for the label-free optical transduction of chemical reactions. Recently, we have found that certain complexes formed by arginine-rich cell-penetrating peptides (CPPs) and amphiphilic counteranions can act as synergistic anion carriers in lipid bilayer membranes. Application of this discovery to rapid and reversible cytosolic CPP delivery has been described (Futaki, S.; et al. ACS Chem. Biol. 2006, 1, 299). Here, we report the complementary use of polyarginine (pR)-counteranion complexes as general optical transducers of chemical reactions. Counterion screening revealed dodecyl phosphate (DP) as an ideal pR activator. Carboxyfluorescein (CF)-loaded vesicles with a shelf life of 3.5 years served best for the detection of fluorogenic CF release by pR-DP complexes with the naked eye. Inactivation of pR-DP complexes by counterion exchange with hyaluronan (HA) caused no CF emission, while HA removal by hyaluronidase (HAase) did. pR-DP complexes were further compatible with the optical detection of HA immobilization on solid support as well as inhibitor screening for HAase (cromolyn, heparin) with and without substrate immobilization. Controls concerning binary ATP/ADP discrimination for naked-eye kinase detection are mentioned to delineate scope but also limitations of this simple and quite universal method.
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PMID: 16984184
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MIYATAKE, Tomohiro, NISHIHARA, Masamichi, MATILE, Stefan. A Cost-Effective Method for the Optical Transduction of Chemical Reactions. Application to Hyaluronidase Inhibitor Screening with Polyarginine-Counteranion Complexes in Lipid Bilayers. In: Journal of the American Chemical Society, 2006, vol. 128, n° 38, p. 12420-12421. doi: 10.1021/ja064732o https://archive-ouverte.unige.ch/unige:6918

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Deposited on : 2010-06-18

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