FLUOROGENIC SUBSTRATES FOR Β-D-Galactosidases AND PHOSPHATASES DERIVED FROM FLUORESCEIN (3, 6-Dihydroxyfluoran) AND ITS MONOMETHYL ETHER

Rotman, Boris; Zderic, John A.; Edelstein, M.P.

doi:10.1073/pnas.50.1.1

Cited by 127 publications

(69 citation statements)

References 4 publications

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“…Elution was performed at 30°C and 1 ml/min with a linear gradient rising within 15 min from 7 % to 63 % acetonitrile in 0.1 M sodium phosphate pH 3, proved useful in the separation of kinins [13]. Detection wavelength was 273 nm where FDG has an absorbance maximum [14] which was also found at 272-274 nm in FMG and fluorescein under HPLC conditions. FDG was eluted at 8.5 min, FMG at 10.7 min, and fluorescein at 13.7 min.…”

Section: Quantitative Hplc Of Fdg Fmg and Fluoresceinmentioning

confidence: 99%

No intermediate channelling in stepwise hydrolysis of fluorescein di‐β‐D‐galactoside by β‐galactosidase

Fiedler¹,

Hinz²

1994

European Journal of Biochemistry

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In the enzymic hydrolysis of biopolymers such as polypeptides, polysaccharides or nucleic acids, two extreme patterns of action are conceivable [l]. The enzyme, after forming an enzyme-substrate complex, may begin to catalyze hydrolysis of mult.iple bonds in what has been described as zipper fashion until it comes to the end of the chain. Alternatively, the hydrolysis of only one bond per effective encounter is catalyzed, the classical random action. Between these extremes is the case of 'multiple attack', where several bonds are hydrolyzed before the enzyme-substrate complex dissociates. Examples for the hydrolysis of more than one bond per enzyme-substrate encounter seem to be relatively rare. Multiple attack has been reported in the hydrolysis of starch by a-and P-amylases [l, 21. 'Processivity' (as it is called there) is observed in enzymes processing the highly charged DNAs or RNAs and might be a feature of their polymerase function, retained e.g. in the partially processive hydrolysis by the RNase H activity of retroviral reverse transcriptase [3]. Processive hydrolysis is also found with ribonuclease I1 [4] and exonuclease I from Escherichiu coli and some further exonucleases As to cases where only a few specific bonds are to be hydrolyzed in a substrate molecule, partial processive cleavage was described in the hydrolysis of polynucleotides with two EcoRI sites by EcoRI endonuclease [6, 71. We have obtained evidence that in the release of the kallidin decapeptide from bovine kininogen by tissue kallikrein the two peptide bonds hydrolyzed may be cleaved at a single enzyme-substrate encounter [8, 91. Therefore, we were highly interested in a recent report [lo] stating that in the hydrolysis of fluorescein di-P-galactoside (FDG) by P-galactosidase intermediate channelling does occur.

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Section: Quantitative Hplc Of Fdg Fmg and Fluoresceinmentioning

confidence: 99%

No intermediate channelling in stepwise hydrolysis of fluorescein di‐β‐D‐galactoside by β‐galactosidase

Fiedler¹,

Hinz²

1994

European Journal of Biochemistry

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“…12b, 21 This strategy has been used to develop reporters for various target enzymes, such as glycosidases, 12c,21 phosphatases, 11a and peptidases. 8c,22 One example is the case of β-galactosidase, an enzyme that is widely used in reporter gene assay.…”

Section: ·1 Masking Groupmentioning

confidence: 99%

Evaluation of Enzymatic Activities in Living Systems with Small-molecular Fluorescent Substrate Probes

Komatsu

Urano

2015

ANAL. SCI.

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“…4). [71][72][73] Substrate 9 is hydrolysed in a sequential manner to release fluorescein whose fluorescence can be monitored at 514 nm. The intermediate, fluorescein mono-b-D-galactopyranoside (FMG) 10, although a good substrate for bGal, has a strong fluorescence and absorbance at the wavelength used for detection of fluorescein thereby hampering its utility.…”

Section: Fluoresceinmentioning

confidence: 99%

Recent advances in the development of synthetic chemical probes for glycosidase enzymes

2015

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The emergence of synthetic glycoconjugates as chemical probes for the detection of glycosidase enzymes has resulted in the development of a range of useful chemical tools with applications in glycobiology, biotechnology, medical and industrial research. Critical to the function of these probes is the preparation of substrates containing a glycosidic linkage that when activated by a specific enzyme or group of enzymes, irreversibly releases a reporter molecule that can be detected. Starting from the earliest examples of colourimetric probes, increasingly sensitive and sophisticated substrates have been reported. In this review we present an overview of the recent advances in this field, covering an array of strategies including chromogenic and fluorogenic substrates, lanthanide complexes, gels and nanoparticles. The applications of these substrates for the detection of various glycosidases and the scope and limitations for each approach are discussed.

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FLUOROGENIC SUBSTRATES FOR Β-D-Galactosidases AND PHOSPHATASES DERIVED FROM FLUORESCEIN (3, 6-Dihydroxyfluoran) AND ITS MONOMETHYL ETHER

Cited by 127 publications

References 4 publications

No intermediate channelling in stepwise hydrolysis of fluorescein di‐β‐D‐galactoside by β‐galactosidase

No intermediate channelling in stepwise hydrolysis of fluorescein di‐β‐D‐galactoside by β‐galactosidase

Evaluation of Enzymatic Activities in Living Systems with Small-molecular Fluorescent Substrate Probes

Recent advances in the development of synthetic chemical probes for glycosidase enzymes

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