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Two-dimensional electrophoresis of cereal prolamins: applications to biochemical and genetic analyses.
Electrophoresis. 1988 Nov; 9(11):727-37.E

Abstract

Three complementary two-dimensional systems for the analysis of cereal prolamins are described. These are electrophoresis at pH 3.1 followed by electrophoresis at pH 9.2, isoelectric focusing (IEF) followed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and SDS-PAGE under non-reducing conditions followed by SDS-PAGE with reduction. They together give information on the pIs, Mrs and charge properties of the individual prolamin components, and on their interactions to form oligomers stabilized by inter-chain disulphide bonds. The three systems are then compared for their effectiveness in analysing unreduced prolamin I fractions from wheat and rye, and the IEF/SDS-PAGE system for analysing reduced and pyridylethylated prolamin fractions from all the major cereals. Finally, applications of the systems in biochemical and genetic studies are discussed and illustrated with three examples: analysis of the structural relationships of the S-rich prolamins (B hordeins and gamma-type hordeins) of barley, determination of the interactions of wheat and rye prolamin subunits in a 2RS/2BL translocation line, and the mapping of genes for alpha-type prolamins in the wild grass Haynaldia villosa.

Authors+Show Affiliations

Shewry PR
AFRC Institute of Arable Crops Research, Rothamsted Experimental Station, Harpenden, Herts, UK.
Parmar S
No affiliation info available
Field JM
No affiliation info available

MeSH

Chromosome MappingEdible GrainElectrophoresis, Gel, Two-DimensionalMolecular StructurePlant ProteinsPolymorphism, GeneticProlaminsSecaleTranslocation, GeneticTriticum

Pub Type(s)

Journal Article

Language

eng

PubMed ID

3250875

Citation

Shewry, P R., et al. "Two-dimensional Electrophoresis of Cereal Prolamins: Applications to Biochemical and Genetic Analyses." Electrophoresis, vol. 9, no. 11, 1988, pp. 727-37.
Shewry PR, Parmar S, Field JM. Two-dimensional electrophoresis of cereal prolamins: applications to biochemical and genetic analyses. Electrophoresis. 1988;9(11):727-37.
Shewry, P. R., Parmar, S., & Field, J. M. (1988). Two-dimensional electrophoresis of cereal prolamins: applications to biochemical and genetic analyses. Electrophoresis, 9(11), 727-37.
Shewry PR, Parmar S, Field JM. Two-dimensional Electrophoresis of Cereal Prolamins: Applications to Biochemical and Genetic Analyses. Electrophoresis. 1988;9(11):727-37. PubMed PMID: 3250875.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Two-dimensional electrophoresis of cereal prolamins: applications to biochemical and genetic analyses. AU - Shewry,P R, AU - Parmar,S, AU - Field,J M, PY - 1988/11/1/pubmed PY - 1988/11/1/medline PY - 1988/11/1/entrez SP - 727 EP - 37 JF - Electrophoresis JO - Electrophoresis VL - 9 IS - 11 N2 - Three complementary two-dimensional systems for the analysis of cereal prolamins are described. These are electrophoresis at pH 3.1 followed by electrophoresis at pH 9.2, isoelectric focusing (IEF) followed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and SDS-PAGE under non-reducing conditions followed by SDS-PAGE with reduction. They together give information on the pIs, Mrs and charge properties of the individual prolamin components, and on their interactions to form oligomers stabilized by inter-chain disulphide bonds. The three systems are then compared for their effectiveness in analysing unreduced prolamin I fractions from wheat and rye, and the IEF/SDS-PAGE system for analysing reduced and pyridylethylated prolamin fractions from all the major cereals. Finally, applications of the systems in biochemical and genetic studies are discussed and illustrated with three examples: analysis of the structural relationships of the S-rich prolamins (B hordeins and gamma-type hordeins) of barley, determination of the interactions of wheat and rye prolamin subunits in a 2RS/2BL translocation line, and the mapping of genes for alpha-type prolamins in the wild grass Haynaldia villosa. SN - 0173-0835 UR - https://www.unboundmedicine.com/medline/citation/3250875/Two_dimensional_electrophoresis_of_cereal_prolamins:_applications_to_biochemical_and_genetic_analyses_ DB - PRIME DP - Unbound Medicine ER -