Proteomic analysis of amphiphilic proteins of hexaploid wheat kernels.
Wheat proteins and specially gluten proteins have been well studied and are closely associated with baking products. Amphiphilic proteins (proteins that are soluble using nonionic detergent Triton X-114 ) also play an important role in wheat quality. Some of them, like puroindolines, are lipid binding proteins, and are strongly linked to dough foaming properties and to fine crumb texture. However many amphiphilic proteins are still unknown and both their physiological and technological functions remain to be analysed. In order to explore these proteins, proteomic analysis was carried out using 81 F9 lines, progeny obtained from an interspecific cross "W7984"x"Opata", and already used to built a map of more than 2000 molecular markers (International Triticeae Mapping Initiative, ITMImap). Two-dimensional electrophoresis (immobilized pH gradient (pH 6-11)x sodium dodecyl sulfate-polyacrylamide gel electrophoresis) was performed on amphiphilic proteins with three to five replicates for each line. Silver stained gels were analysed using Melanie 3 software. Genetic determinism was carried out on 170 spots segregating between the two parental hexaploïd wheats. Many of these spots were mapped on different chromosomes of the ITMImap. Spots of interest were identified using matrix-assisted laser desorption/ionization-time of flight and some of them were partly sequenced using electrospray ionization-tandem mass spectrometry. This proteomic approach provided some very useful information about some proteic components linked to bread wheat quality and particularly to kernel hardness.
UMR-ASP-UBP, Clermont Ferrand, France.
Nardjis
N
Amiour
Marielle
M
Merlino
Philippe
P
Leroy
Gérard
G
Branlard
D002874:Chromosome Mapping; D003433:Crosses, Genetic; D003902:Detergents / Q000494:pharmacology; D015180:Electrophoresis, Gel, Two-Dimensional; D017343:Genes, Plant; D013058:Mass Spectrometry; D008957:Models, Genetic; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000494:pharmacology; D011506:Proteins / Q000737:chemistry* / Q000235:genetics*; D020543:Proteome / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014908:Triticum / Q000235:genetics* / Q000378:metabolism*
D003902:Detergents; D011506:Proteins; D020543:Proteome; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40