A proteomic approach to verify in vivo expression of a novel gamma-gliadin containing an extra cysteine residue.
Gliadins and glutenins are the main protein fractions present in wheat gluten. They are responsible for technological and nutritional quality of wheat based products. In particular, glutenins are mainly responsible for dough visco-elastic properties, whereas gliadins confer extensibility to dough and are the most important factor triggering celiac disease, the major human intolerance to gluten. Gliadins are monomeric proteins, whereas glutenins are polymers stabilized by disulfide bonds. Although they have distinctive structural characteristics, it is possible that some gliadins become part of the glutenin fraction because of mutations that affect cysteine number and distribution. Here, we provide evidence that a naturally mutated gamma-gliadin with an extra cysteine residue is incorporated into the polymeric fraction. This goal was achieved using an integrated approach involving heterologous expression, 2-DE, RP-HPLC and MS.
Ferrante
Paola
P
Dipartimento di Agrobiologia e Agrochimica, Università degli Studi della Tuscia, Viterbo, Italy.
Masci
Stefania
S
D'Ovidio
Renato
R
Lafiandra
Domenico
D
Volpi
Chiara
C
Mattei
Benedetta
B
D000595:Amino Acid Sequence; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D017124:Conserved Sequence; D003553:Cystine / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005433:Flour; D015870:Gene Expression; D017343:Genes, Plant; D005822:Genetic Vectors; D005903:Gliadin / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D005983:Glutens / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D002479:Inclusion Bodies / Q000737:chemistry; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000378:metabolism; D010449:Peptide Mapping; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014908:Triticum / Q000737:chemistry / Q000235:genetics
D003553:Cystine; D005983:Glutens; D005903:Gliadin; D010447:Peptide Hydrolases