A fluorescent-based HPLC assay for quantification of cysteine and cysteamine adducts in Escherichia coli-derived proteins

Soriano, Brian D.; Tam, Lei-Ting; Lu, Hsieng S.; Valladares, Violeta G.

doi:10.1016/j.jchromb.2011.11.011

Cited by 14 publications

(18 citation statements)

References 27 publications

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“…Therefore, the assessment of cellular and plasma levels of therapeutically administered cysteamine is of utmost importance for disease management. Thiol compounds have been detected by a variety of reagents and separation techniques such as HPLC, GC, CE and mass spectrometry between others as it has been published (Bayle et al 2004, Carlucci and Tabucchi 2009, Dohil et al 2012, Kataoka et al 1994, McMenamin et al 2009, Soriano et al 2012). It is uncertain whether cysteamine plasma levels reflect cysteamine tissue levels.…”

Section: Caveatsmentioning

confidence: 99%

Cysteamine revisited: repair of arginine to cysteine mutations

Gallego

Hannibal

Häberle

et al. 2017

J of Inher Metab Disea

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Cysteamine is a small aminothiol endogenously derived from coenzyme A degradation. For some decades, synthetic cysteamine has been employed for the treatment of cystinosis, and new uses of the drug continue to emerge. In this review, we discuss the role of cysteamine in cellular and extracellular homeostasis and focus on the potential use of aminothiols to reconstitute the function of proteins harboring arginine (Arg) to cysteine (Cys) mutations, via repair of the Cys residue into a moiety that introduces an amino group, as seen in basic amino acid residues Lys and Arg. Cysteamine has been utilized in vitro and ex vivo in four different genetic disorders, and thus provides “proof of principle” that aminothiols can modify Cys residues. Other aminothiols such as mercaptoethylguanidine (MEG) with closer structural resemblance to the guanidinium moiety of Arg are under examination for their predicted enhanced capacity to reconstitute loss of function. Although the use of aminothiols holds clinical potential, more studies are required to refine specificity and treatment design. The efficacy of aminothiols to target proteins may vary substantially depending on their specific extracellular and intracellular locations. Redox potential, pH and specific aminothiol abundance in each physiological compartment are expected to influence the reactivity and turnover of cysteamine and analogous drugs. Upcoming research will require the use of suitable cell and animal models featuring Arg to Cys mutations. Since Arg to Cys changes comprise in general about 8% of missense mutations, repair of this specific mutation may provide promising avenues for many genetic diseases.

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Section: Caveatsmentioning

confidence: 99%

Cysteamine revisited: repair of arginine to cysteine mutations

Gallego

Hannibal

Häberle

et al. 2017

J of Inher Metab Disea

View full text Add to dashboard Cite

show abstract

“…These fluorescent reagents were reported previously to be useful for the fluorometric detection of proteins, peptides, amino-and imino-acid [24][25][26]. A 10 µL extract sample of each treatment was added to 100 µL of 0.266 M NaHCO 3 and 80 µL of 4.75 mg mL -1 NBD-Cl/MeOH.…”

Section: Derivatization By Fluorescent Reagentsmentioning

confidence: 99%

“…Previous studies reported that the NBD-derivatized proteins and peptides have been separated using molecular exclusion chromatography [29] and reversed phase chromatography [26]. The phytoplankton extracts in this study would contain biological soluble molecules that have more than 5 kDa of molecular weight.…”

Section: Selection Of Fluorescent Reagentmentioning

confidence: 99%

“…Fluorescent-based HPLC assay is a quick, easy, efficient and powerful method for the quantitative measurement of cellular macromolecules in organisms [22][23][24][25][26][27]. This method has been used for the determination of cysteine and cysteamine adducts in Escherichia coli-derived proteins [26], thiol compounds in biological samples [22], human erythrocytic glutathione [25], domoic acid (DA) in mussels (Mytilus edulis) [24], peptides in tissue and plasma samples [28] and amines and proteins [29] using different fluorescent reagents as the derivatizer.…”

Section: Introductionmentioning

confidence: 99%

“…This method has been used for the determination of cysteine and cysteamine adducts in Escherichia coli-derived proteins [26], thiol compounds in biological samples [22], human erythrocytic glutathione [25], domoic acid (DA) in mussels (Mytilus edulis) [24], peptides in tissue and plasma samples [28] and amines and proteins [29] using different fluorescent reagents as the derivatizer. The HPLC method has also been used for the determination of pigments in phytoplankton in order to measure marine productivity in relation of Fe status in seawater [30,31].…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

A Fluorescent-Based HPLC Assay Using 4-Chloro-7-nitrobenzo-2-oxa-1, 3-diazole as Derivatization Agent for the Determination of Iron Bioavailability to Red Tide Phytoplankton

et al. 2014

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A new fluorescent-based high performance liquid chromatography (HPLC) assay using 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole (NBD-C1) was employed to determine iron (Fe) bioavailability to red tide phytoplankton in seawater. After growing four red tide species (Prymnesium parvum, Heterosigma akashiwo, Eutreptiella gymnastica, and Oltmannsiellopsis viridis) in f/2 artificial seawater under different Fe conditions, soluble extracts of the phytoplankton were derivatized using different fluorescent reagents (NBD-C1, 4-fluoro-7-nitrobenzo-2-oxa-1,3-diazole; NBD-F, fluorescamine, and ortho-phthalaldehyde; OPA) followed by HPLC assay. Among the four fluorescent reagents, NBD-C1 was most effective for derivatizing the phytoplankton extracts which would consist of proteins and peptides. HPLC chromatograms of the NBDderivatized extracts showed gradual changes (decrease/increase) of six peaks for different Fe conditions. Four of the peaks decreased, while two peaks increased with the increase of Fe concentrations in the culture medium. Considering the consistency and sensitivity of chromatogram peaks E and A to different Fe, phosphate and nitrate conditions for all phytoplankton studied, the ratio of these two peaks (I E/A ) has been proposed as the indicator of Fe bioavailability to red tide phytoplankton.

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Enantioseparation of 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate tagged amino acids and other zwitterionic compounds on cinchona-based chiral stationary phases

2013

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The fluorescent tag 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC; AccQ Fluor reagent kit from Waters) is a commercial N-terminal label for proteinogenic amino acids (AAs), designed for reversed-phase separation and quantification of the AA racemates. The applicability of AQC-tagged AAs and AA-type zwitterionic compounds was tested for enantiomer separation on the tert-butyl carbamate modified quinine and quinidine based chiral stationary phases, QN-AX and QD-AX employing polar-organic elution conditions. The investigated test analytes included the enantiomers of the positional isomers of isoleucine (Ile), threonine, homoserine, and 4-hydroxyproline. Furthermore, β-AAs, cyclic, and heterocyclic AAs including trans-2-amino-cyclohexane carboxylic acid and trans-2-aminocyclohexyl sulfonic acid, phenylalanine derivatives substituted with halides with increasing electronegativity and 3,4-dihydroxyphenylalanine, cysteine-related derivatives including homocysteic acid, methionine sulfone, cysteine-S-acetic acid, and cysteine-S-acetamide as well as a small range of aminophosphonic acids were enantioseparated. A mechanistic interaction study of AQC-AAs in comparison with fluoresceine isothiocyanate-labeled AAs was performed. The chiral and chemoselective recognition processes involved in enantiomer separation and retention was systematically discussed. Special emphasis was set on the influential factors exhibited by the chemistry, branching position, and spatial properties of the investigated zwitterionic analytes. The general interest to separate and distinguish between different types of branched-chained AAs and metabolic side products thereof lies in the toxicity of some of these compounds, which makes for instance allo-Ile an attractive candidate in disease-related biomarker research.

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A fluorescent-based HPLC assay for quantification of cysteine and cysteamine adducts in Escherichia coli-derived proteins

Cited by 14 publications

References 27 publications

Cysteamine revisited: repair of arginine to cysteine mutations

Cysteamine revisited: repair of arginine to cysteine mutations

A Fluorescent-Based HPLC Assay Using 4-Chloro-7-nitrobenzo-2-oxa-1, 3-diazole as Derivatization Agent for the Determination of Iron Bioavailability to Red Tide Phytoplankton

Enantioseparation of 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate tagged amino acids and other zwitterionic compounds on cinchona-based chiral stationary phases

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