Novel Chromogenic Substrates for the Rate-Assay of N-Acetyl-β-D-glucosaminidase: Resorufinyl- and Resazurinyl-N-acetyl-β-D-glucosaminides

Kasai, Kouichi; Yamaji, Nobuyuki

doi:10.2116/analsci.8.161

Cited by 5 publications

(3 citation statements)

References 10 publications

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“…On the basis of this mechanism, various O-alkyl resorufin derivatives have been synthesized as turn-on fluorogenic substrates of enzymes. [16][17][18][19][20] For example, Chang and co-workers reported that O-3-aminopropyl resorufin derivatives act as turn-on fluorogenic substrates for monoamine oxidases (MAOs). 21 The MAO probes are converted to resorufin via enzymatic oxidation of the amine group followed by b-elimination, as shown in Scheme 1.…”

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confidence: 99%

Iron complex-based fluorescent probes for intracellular hydrogen peroxide detection

2013

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confidence: 99%

Iron complex-based fluorescent probes for intracellular hydrogen peroxide detection

2013

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“…Enzymatic consumption and release of 2 have been demonstrated to function as useful indicator reactions for spectrophotometric and fluorometric analyses, since the dye exhibits strong absorption (l max 571 nm, e 4-7ϫ10 4 ) and emission (excitation maximum at 563 nm and emission maximum at 587 nm at pH 7.4) at wavelengthsϾ550 nm, where potential interference in analysis of colored or turbid serum components can be avoided. [25][26][27][28][29][30][31][32][33][34][35] These observations imply that the transformation of 1 to 2 as an indicator reaction for glucose analysis using only GOD takes full advantage of 2 as an analytically useful dye.…”

Section: Resultsmentioning

confidence: 99%

“…As mentioned above, several analytical methods with spectrophotometry or fluorometry have been designed based on enzymatic release of 2 from its derivatives such as 1. [25][26][27][28][29][30][31][32][33][34][35] However, to our knowledge, generation of 2 from 1 has been used as an indicator reaction only for studies of esterase activity of cytosolic aldehyde dehydrogenase and chymotrypsin. [20][21][22][23] Thus, this is the first report to show that transformation of 1 to 2 is induced by H 2 O 2 .…”

Section: Resultsmentioning

confidence: 99%

Hydrogen Peroxide-Induced Deacetylation of Acetyl Resorufin as a Novel Indicator Reaction for Fluorometric Detection of Glucose Using Only Glucose Oxidase.

Maeda

Matsu-Ura

Nishida

et al. 2001

CHEMICAL & PHARMACEUTICAL BULLETIN

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Spectrophotometric or fluorometric methods with glucose oxidase (GOD)-peroxidase (POD)-chromogen(s) systems are well known as useful tools for determination of glucose. [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15] Indicator reactions in these methods are based on oxidative formation of a colored or fluorescent substance from a chromogen(s) with H 2 O 2 , generated through GODcatalyzed oxidation of glucose, in the presence of POD. These color reactions are well-characterized chemical processes. However, the POD-dependent indicator reactions are known to be inevitably disturbed by electron donors present in biological samples such as ascorbic acid, uric acid, and bilirubin.3) Major processes of interference by these compounds are as follows: 1) reduction of oxidatively formed colored or fluorescent substance to its original chromogen(s); and 2) competition with a chromogen in reduction of H 2 O 2 . Several methodologies have been developed for elimination of such interference in glucose determination with GOD-POD-chromogen(s) systems.3) However, the most straightforward approach to glucose determination free from such interference is design of a novel indicator reaction without recourse to redox reactions coupled with H 2 O 2 and POD. To our knowledge, only a few methods with non-redox color reactions for glucose determination using only GOD have been reported. These methods utilize transformation of a vanadium(V), 16) titanium(IV) 17) or dinuclear iron(III) 18) complex to its adduct with H 2 O 2 accompanied by a bathochromic shift as a spectrophotometric indicator reaction. As expected, it was demonstrated that glucose determination with vanadium(V) and titanium(IV) complexes was not affected by various substances usually present in serum or added to test solution. 16,17) However, spectrophotometric or fluorometric determination of glucose with high accuracy should use formation of a colored or fluorescent substance from a chromogen rather than a color change of a dye as an indicator reaction.Recently, resorufin (2) was shown to be able to reoxidize a reduced form of GOD at 37°C, being transformed to a colorless dihydro derivative, although the reductive bleaching of the dye is of no use for GOD-based determination of glucose as an indicator reaction. 19) This finding prompted us to examine how resorufin derivatives such as acetyl resorufin (1) would behave in GOD-catalyzed oxidation of glucose, and it was found that 1 behaves in a manner different from 2 in the enzymatic reaction. Here, we report that deacetylation of non-fluorescent 1 to fluorescent 2 is induced by H 2 O 2 generated in GOD-catalyzed oxidation of glucose, and the transformation is promising as an indicator reaction for fluorometric determination of glucose without significant effects of ascorbic acid, uric acid, or bilirubin. ExperimentalMaterials GOD from Aspergillus niger (EC 1.1.3.4) and glucose were used as supplied from Wako Pure Chemical Industries, Ltd. Acetyl resorufin (1) [20][21][22][23] was prepared by reaction of resorufin sod...

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A potent mechanism-inspired O-GlcNAcase inhibitor that blocks phosphorylation of tau in vivo

et al. 2008

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Pathological hyperphosphorylation of the microtubule-associated protein tau is characteristic of Alzheimer's disease (AD) and the associated tauopathies. The reciprocal relationship between phosphorylation and O-GlcNAc modification of tau and reductions in O-GlcNAc levels on tau in AD brain offers motivation for the generation of potent and selective inhibitors that can effectively enhance O-GlcNAc in vertebrate brain. We describe the rational design and synthesis of such an inhibitor (thiamet-G, K(i) = 21 nM; 1) of human O-GlcNAcase. Thiamet-G decreased phosphorylation of tau in PC-12 cells at pathologically relevant sites including Thr231 and Ser396. Thiamet-G also efficiently reduced phosphorylation of tau at Thr231, Ser396 and Ser422 in both rat cortex and hippocampus, which reveals the rapid and dynamic relationship between O-GlcNAc and phosphorylation of tau in vivo. We anticipate that thiamet-G will find wide use in probing the functional role of O-GlcNAc in vertebrate brain, and it may also offer a route to blocking pathological hyperphosphorylation of tau in AD.

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Novel Chromogenic Substrates for the Rate-Assay of N-Acetyl-β-D-glucosaminidase: Resorufinyl- and Resazurinyl-N-acetyl-β-D-glucosaminides

Cited by 5 publications

References 10 publications

Iron complex-based fluorescent probes for intracellular hydrogen peroxide detection

Iron complex-based fluorescent probes for intracellular hydrogen peroxide detection

Hydrogen Peroxide-Induced Deacetylation of Acetyl Resorufin as a Novel Indicator Reaction for Fluorometric Detection of Glucose Using Only Glucose Oxidase.

A potent mechanism-inspired O-GlcNAcase inhibitor that blocks phosphorylation of tau in vivo

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