title: Glutamine-binding protein from Escherichia coli specifically binds a wheat gliadin peptide. 2. Resonance energy transfer studies suggest a new sensing approach for an easy detection of wheat gliadin.
abstract: In this work is presented the first attempt to develop a fluorescence assay for detection of traces of gluten in food by utilizing the recombinant glutamine-binding protein (GlnBP) from E. coli. We found that GlnBP specifically binds the sequence of amino acids present both in gliadin and other prolamines classified as toxic for celiac patients. Affinity chromatography experiments together with mass spectrometry experiments demonstrated that GlnBP can bind the following amino acid sequence XXQPQPQQQQQQQQQQQQL. Sequence alignment experiments pointed out that this sequence is exclusively representative of the gliadin and the other prolamines considered toxic for celiac patients. These findings suggest the development of a competitive resonance energy transfer (RET) assay for an easy and rapid detection of this sequence in raw and cooked food.
authors.affiliation: Institute of Protein Biochemistry, C.N.R., Via Pietro Castellino, 111 80131, Naples, Italy.
authors.forename: Maria
authors.initials: M
authors.lastname: Staiano
authors.forename: Viviana
authors.initials: V
authors.lastname: Scognamiglio
authors.forename: Gianfranco
authors.initials: G
authors.lastname: Mamone
authors.forename: Mauro
authors.initials: M
authors.lastname: Rossi
authors.forename: Antonietta
authors.initials: A
authors.lastname: Parracino
authors.forename: Mose'
authors.initials: M
authors.lastname: Rossi
authors.forename: Sabato
authors.initials: S
authors.lastname: D'Auria
mesh_terms: D000595:Amino Acid Sequence; D002352:Carrier Proteins / Q000378:metabolism*; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000235:genetics*; D005504:Food Analysis / Q000379:methods*; D005903:Gliadin / Q000302:isolation & purification* / Q000378:metabolism; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D011485:Protein Binding; D016415:Sequence Alignment; D013050:Spectrometry, Fluorescence; D014908:Triticum / Q000235:genetics*
chemical_list: D002352:Carrier Proteins; C037853:glutamine transport proteins; D005903:Gliadin