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.
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.
Staiano
Maria
M
Institute of Protein Biochemistry, C.N.R., Via Pietro Castellino, 111 80131, Naples, Italy.
Scognamiglio
Viviana
V
Mamone
Gianfranco
G
Rossi
Mauro
M
Parracino
Antonietta
A
Rossi
Mose'
M
D'Auria
Sabato
S
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*
D002352:Carrier Proteins; C037853:glutamine transport proteins; D005903:Gliadin