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Volumn 60, Issue 34, 2004, Pages 7267-7275

FRET induced by an 'allosteric' cycloaddition reaction regulated with exogenous inhibitor and effectors

Author keywords

Allosteric catalysis; ELISA; F rster resonance energy transfer; Husigen cycloaddition; Lead; Metal ion detection; Signal amplification; T stack; Zinc

Indexed keywords

ASCORBIC ACID; COPPER; COPPER SULFATE; EDETIC ACID; LEAD; ORGANOMETALLIC COMPOUND; RIBOZYME; TRANSITION ELEMENT; ZINC ION;

EID: 3342903448     PISSN: 00404020     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.tet.2004.06.079     Document Type: Conference Paper
Times cited : (47)

References (48)
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    • Ascorbate acid activity might be regulated by a saccharide receptor
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    • Although the Cu(I) catalyzed Huisgen cycloaddition is highly efficient, it is after all a bimolecular reaction. The reactants concentrations under this study were ∼1 mM, which is 200 times less than reported in the original study (Ref. 15b). Under such conditions the reaction was not completed in 18 h. However, the product abundance is sufficient enough to be monitored by HPLC and have a substantial modulating effect on the fluorescence
    • Although the Cu(I) catalyzed Huisgen cycloaddition is highly efficient, it is after all a bimolecular reaction. The reactants concentrations under this study were ∼1 mM, which is 200 times less than reported in the original study (Ref. 15b). Under such conditions the reaction was not completed in 18 h. However, the product abundance is sufficient enough to be monitored by HPLC and have a substantial modulating effect on the fluorescence
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    • In a separate experiment, bromide anion was also shown by fluorescence to slightly slow the reaction down
    • In a separate experiment, bromide anion was also shown by fluorescence to slightly slow the reaction down
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    • For general practice FRET signal should be denoted as the intensity ratio between acceptor and donor emissions (when donor is excited). However in our case, the donor emission was usually too high to measure, especially when the metal ion effector concentration was low. Therefore, the intensity at 500 nm was used as an alternative measure. That might account for the slight difference between fluorescence and HPLC analysis patterns. HPLC analyses were conducted for other reactions sets as well and they generally match well with the related fluorescence profiles
    • For general practice FRET signal should be denoted as the intensity ratio between acceptor and donor emissions (when donor is excited). However in our case, the donor emission was usually too high to measure, especially when the metal ion effector concentration was low. Therefore, the intensity at 500 nm was used as an alternative measure. That might account for the slight difference between fluorescence and HPLC analysis patterns. HPLC analyses were conducted for other reactions sets as well and they generally match well with the related fluorescence profiles


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