Quantification of riboflavin, flavin mononucleotide, and flavin adenine dinucleotide in mammalian model cells by CE with LED‐induced fluorescence detection

  title={Quantification of riboflavin, flavin mononucleotide, and flavin adenine dinucleotide in mammalian model cells by CE with LED‐induced fluorescence detection},
  author={Jens H{\"u}hner and {\'A}lvaro Ingl{\'e}s-Prieto and Christian Neus{\"u}ss and Michael L{\"a}mmerhofer and Harald Janovjak},
Cultured mammalian cells essential are model systems in basic biology research, production platforms of proteins for medical use, and testbeds in synthetic biology. Flavin cofactors, in particular flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), are critical for cellular redox reactions and sense light in naturally occurring photoreceptors and optogenetic tools. Here, we quantified flavin contents of commonly used mammalian cell lines. We first compared three procedures for… 

Bioenergetic Alterations of Metabolic Redox Coenzymes as NADH, FAD and FMN by Means of Fluorescence Lifetime Imaging Techniques

The FLIRR approach was extended and new results were presented, which includes FLIM data of the various enzymes, such as NAD(P)H, FAD, as well as flavin mononucleotide (FMN), which used to calculate the fluorescence lifetime induced redox ratio (FLIRR), which was found to be highest for tumor and normal cells.

Enhanced small green fluorescent proteins as a multisensing platform for biosensor development

The miniGFPs proteins exhibited improved photochemical properties compared to other flavin-binding FPs enabling long-term live cell imaging and can serve as a multisensing platform for fluorescence biosensor development for in vitro and in-cell applications.

Self-assembly of flavin mononucleotide and a cationic polythiophene in aqueous media: spectroscopic studies and sensing applications

The synergistic effect of non-covalent interactions containing electrostatic, hydrophobic and π–π interactions is proposed to account for the formation of PMT-1/FMN nano-superstructures, which features wide detection range, fast response, high sensitivity and selectivity.

Extracellular pH affects the fluorescence lifetimes of metabolic co-factors

Autofluorescence measurements of the metabolic cofactors NADH and flavin adenine dinucleotide provide a label-free method to quantify cellular metabolism and changes in endogenous fluorescence lifetimes with extracellular pH are mostly due to indirect changes within the cell rather than direct pH quenching of the endogenous molecules.

Fluorescence intensity and lifetime redox ratios detect metabolic perturbations in T cells.

While both the fluorescence lifetime and intensity redox ratios resolve metabolic perturbations in T cells, the endpoints are influenced by different metabolic processes.

Fluorescence intensity and lifetime redox ratios detect metabolic perturbations in T cells

: The auto-fluorescent coenzymes reduced nicotinamide dinucleotide (NADH) and oxidized flavin adenine dinucleotide (FAD) allow label-free detection of cellular metabolism. The optical redox ratio,

Identification of Flavin Mononucleotide as a Cell-Active Artificial N6 -Methyladenosine RNA Demethylase.

The first identification of a small-molecule modulator that functions as an artificial m6 A demethylase is reported, which sheds light on the development of powerful small molecules as RNA demethylases and new probes for use in RNA biology.

Crystal structures of FMN‐bound and FMN‐free forms of dihydroorotate dehydrogenase from Trypanosoma brucei

Comparisons of B factors of the protein main chain revealed that binding of FMN decreased flexibility of most of the residues at theFMN‐binding site, but increased flexibility of a lid‐like loop structure over the active center, which was ascribed to a conformational change in an FMN‐contacting residue, Asn195, which induced a rearrangement of a hydrogen‐bond network of the residue comprising the lid.



Quantification of riboflavin, flavin mononucleotide, and flavin adenine dinucleotide in human plasma by capillary electrophoresis and laser-induced fluorescence detection.

Capillary electrophoresis with laser-induced fluorescence detection allows determination of all rib oflavin vitamers far below physiological concentrations, and may become a useful tool for the assessment of riboflavin status in humans.

Separation of flavins and nicotinamide cofactors in Chinese hamster ovary cells by capillary electrophoresis.

The protocols for extraction, separation and quantitation are readily adaptable to normal and cancer cell lines for the analysis of endogenous fluorophores and show wide linear range of quantitation.

Determination of riboflavin, flavin mononucleotide and flavin adenine dinucleotide in biological tissues by capillary zone electrophoresis and laser‐induced fluorescence detection

The separation of riboflavin, flavin mononucleotide and flavin adenine dinucleotide was investigated by capillary zone electrophoresis using laser‐induced fluorescence detection using a systematic approach and was successfully applied to a variety of biological tissues from different animals.

Riboflavin kinase couples TNF receptor 1 to NADPH oxidase

RFK is identified as a previously unrecognized TNF-receptor-1 (TNFR1)-binding protein that physically and functionally couples TNFR1 to NADPH oxidase, and the results suggest that TNF, through the activation of RFK, enhances the incorporation of FAD in NAD PH oxidase enzymes, a critical step for the assembly and activation of NADph oxidase.

Autofluorescence of viable cultured mammalian cells.

  • J. Aubin
  • Biology
    The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society
  • 1979
Comparison of cell spectra with spectra of known cellular metabolites suggested that most, if not all, of the fluorescence arises from intracellular nicotinamide adenine dinucleotide (NADH) and riboflavin and flavin coenzymes.

Laser‐induced fluorescence detection schemes for the analysis of proteins and peptides using capillary electrophoresis

This review article summarizes the works which were performed for protein and peptide analysis via CE‐LIF and noncovalent labelling interactions.