Fluorogenic Label for Biomolecular Imaging

Lavis, Luke D.; Chao, Tzu‐Yuan; Raines, Ronald T.

doi:10.1021/cb600132m

Cited by 190 publications

(219 citation statements)

References 57 publications

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“…Today, the use of fluorescein diacetate and its derivatives as fluorogenic esterase substrates is the basis for a number of cellular assays for viability and long-term tracking (11). More recently, other esterase-active profluorophores have been introduced based on aryl acetate esters possessing higher chemical stability (12).…”

mentioning

confidence: 99%

Ester Bonds in Prodrugs

Lavis

2008

ACS Chem. Biol.

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134

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confidence: 99%

Ester Bonds in Prodrugs

Lavis

2008

ACS Chem. Biol.

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134

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“…3 The widespread use of NMAA active esters as heterobifunctional crosslinkers has resulted in the development of a number of methods 4 for their rapid preparation; however, for applications requiring the free acids the synthetic options are less appealing.…”

Section: Introductionmentioning

confidence: 99%

On the synthesis of N-maleoyl amino acids in aqueous media: cautionary tales for the unwary traveller

Paterson¹,

Jovanović²

2010

Arkivoc

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The high cost or protracted syntheses associated with N-maleoyl amino acid derivatives coupled with a considerable demand from both the life and physical sciences has spurred the search for facile, low-cost routes to these compounds. Herein, we demonstrate that a recently published method purporting to deliver N-maleoyl amino acids by cyclization of maleamic acids in water, instead results in a hydrolysis product, namely the corresponding hemi-maleate salt.

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“…As a dye, we chose a morpholino urea derivative of rhodamine 110 (Rh 110 ) that is bright (ε × Φ = 2.38 × 10 4 M −1 cm −1 ) but has no measurable fluorescence after N-acylation. 10 We installed an ethyl group on the phenolic oxygen of the trimethyl lock because ethyl ethers are especially effective substrates for CYP1A1. 15 The synthetic route to fluorogenic probe 1 is shown in Figure 1.…”

Section: Introductionmentioning

confidence: 99%

A highly sensitive fluorogenic probe for cytochrome P450 activity in live cells

Yatzeck

Lavis

Chao

et al. 2008

Bioorganic & Medicinal Chemistry Letters

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A derivative of rhodamine 110 has been designed and assessed as a probe for cytochrome P450 activity. This probe is the first to utilize a "trimethyl lock" that is triggered by cleavage of an ether bond. In vitro, fluorescence was manifested by the CYP1A1 isozyme with k cat /K M = 8.8 × 10 3 M −1 s −1 and K M = 0.09 µM. In cellulo, the probe revealed the induction of cytochrome P450 activity by the carcinogen 2,3,7,8-tetrachlorodibenzo-p-dioxin, and its repression by the chemoprotectant resveratrol. KeywordsCarcinogen; CYP1A1 Isozyme; Cytochrome P450; Dioxin; Fluorogenic Substrate; Prodrug; Resveratrol; Rhodamine 110; Trimethyl LockThe cytochrome P450 (P450) family of enzymes is responsible for the oxidative metabolism of a wide variety of compounds, including chemotherapeutic agents and environmental toxins. 1,2 The catalytic activity of P450 enzymes controls the rate of xenobiotic metabolism, and can produce undesirable byproducts. 3 Originally, this activity had been assessed by using HPLC or other methods to separate and quantify metabolites. In the 1970's, 7-ethoxycoumarin and 7-ethoxyresorufin were introduced as the first fluorogenic substrates for assays of P450 activity. 4 Although these and other fluorogenic substrates have been used to assay P450 activity in vitro and enable assays in cellulo ,5,6 they suffer from background fluorescence. 7 For example, alkoxycoumarins exhibit moderate fluorescence and are used frequently as fluorophores in peptidase substrates based on Forster resonance energy transfer (FRET). 8 In addition, both 7-ethoxyresorufin and resorufin fluoresce brightly. 6 This problem arises because O-alkylation of the hydroxyl group of fluorophores such as coumarin and resorufin does little to deter the oxygen electrons from participating in the resonance that gives rise to fluorescence.Here, we report on a superior small-molecule probe for assessing P450 activity. Our probe employs the trimethyl lock. 9-11 The trimethyl lock is an o-hydroxycinnamic acid derivative in which severe crowding of three methyl groups induces rapid lactonization to form a hydrocoumarin. 12 In this strategy, the phenolic oxygen of the o-hydroxycinnamic acid is modified to create a functional group that is a substrate for a designated enzyme, and the carboxyl group is condensed with the amino group of a dye. Unmasking of the phenolic oxygen leads to rapid lactonization with concomitant release of the dye. An important attribute of this The human genome contains 27 genes encoding P450 isozymes, along with many pseudogenes. 13 Of these isozymes, cytochrome P450 1A1 (CYP1A1) is known to be especially important in the metabolism of xenobiotics. 14 Unlike most P450 isoforms, which are found primarily in the liver, CYP1A1 is present mainly in the lungs, where it plays an important role in the metabolic activation of chemical carcinogens. 1,15 The lung is a primary site of exposure for inhaled toxins along with carcinogens that can ultimately yield lung carcinomas. 16 CYP1A1 is strongly induced by cigarette smo...

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Fluorogenic Label for Biomolecular Imaging

Cited by 190 publications

References 57 publications

Ester Bonds in Prodrugs

Ester Bonds in Prodrugs

On the synthesis of N-maleoyl amino acids in aqueous media: cautionary tales for the unwary traveller

A highly sensitive fluorogenic probe for cytochrome P450 activity in live cells

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