Diversity of puroindolines as revealed by two-dimensional electrophoresis
Autor: | Thérèse Gaborit, Nardjis Amiour, Stéphane Herbette, Gilberto Igrejas, Gérard Branlard, Didier Marion, Mireille Dardevet |
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Rok vydání: | 2003 |
Předmět: |
0106 biological sciences
Blotting Western Immunoblotting Ion chromatography Mass spectrometry 01 natural sciences Biochemistry Mass Spectrometry Endosperm 0404 agricultural biotechnology Cations Electrophoresis Gel Two-Dimensional Molecular Biology Chromatography High Pressure Liquid Triticum Plant Proteins 2. Zero hunger Gel electrophoresis Polymorphism Genetic Chromatography Spots Chemistry Isoelectric focusing food and beverages 04 agricultural and veterinary sciences Hydrogen-Ion Concentration Chromatography Ion Exchange 040401 food science Electrophoresis Isoelectric point Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Electrophoresis Polyacrylamide Gel 010606 plant biology & botany |
Zdroj: | PROTEOMICS. 3:168-174 |
ISSN: | 1615-9861 1615-9853 |
Popis: | Puroindolines are endosperm lipid binding proteins, which are separated by reversed phase-high-performance liquid chromatography or cation exchange chromatography into two isoforms, puroindoline-a (PIN-a) and puroindoline-b (PIN-b). Being very basic and close in molecular weight, PIN-a and PIN-b have never been separated using conventional isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). A two-dimensional electrophoresis method, linear immobiline pH gradient (IPGxSDS-PAGE), was developed, using 6-11 linear immobiline Dry Strips in the first dimension, which allowed the puroindolines to be focused between isoelectric point 10.5 and 11. Immunoblotting revealed that both PIN-a and PIN-b were each composed of several spots. Two-dimensional patterns from unrelated wheat varieties revealed that several spots can be highlighted among varieties. Matrix-assisted laser desorption/ionization-time of flight spectrometry allowed the majority of the spots revealed in the puroindoline zone to be identified. The two-dimensional IPGxSDS-PAGE of these very basic wheat endosperm proteins, puroindolines and related grain softness proteins should facilitate the identification of the proteins associated with wheat endosperm texture that have a strong effect on milling, dough properties and end-uses of wheats. |
Databáze: | OpenAIRE |
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