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GrainGenes Reference Report: JCS-30-283

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Reference	JCS-30-283
Title	Antibodies of N-terminal peptides of low M(r) subunits of wheat glutenin. II. Detection of subunits encoded by different loci
Journal	Journal of Cereal Science
Year	1999
Volume	30
Pages	283-301
Author	Sissons MJ Hac L Skerritt JH
Abstract	A panel of anti-peptide antibodies specific for each of the different N-terminal sequence types of B- and C-low molecular mass glutenin subunits (LM(r)GS) were utilised in immunoblotting studies to identify the chromosomal location of genes encoding different sequences and to characterise the allelic variation of the encoding loci. The MET-type sequences were predominantly found among the B- subunits, while the alpha- and gamma- sequences predominated in the C- subunits. The quantitatively major SHIPGLERPS sequence was found in both the B- and C- mobility regions. Using either biotypes in the cultivar, Aroona or genetic lines containing double rye chromosome 1 substitutions and thus expressing only single LM(r)GS alleles, the sequences were determined for most of the major polypeptides expressed by each LM(r)GS allele. The LM(r)GS from different genomes encoded different numbers of each sequence type. Furthermore, different polypeptides within a particular block of subunits encoded by a given allele often had differing N-terminal sequences. However, subunits of similar electrophoretic mobilities encoded by different alleles at each locus usually had identical N-terminal sequences, suggesting that they may instead differ in the number of repeats. In Chinese Spring, genes encoding the SHIPGLERPS and METSHIPGL sequence types were predominantly present on chromosomes 1B and 1D, while the related METSRVPGL sequence was only encoded on 1D. In contrast, the METSCIPGL, alpha- and gamma-sequences were encoded on each of chromosomes 1A, 1B and 1D. Several different electrophoretic and immunoblotting approaches using null lines suggested that some of the alpha-type LM(r)GS may also be encoded by group 6 chromosomes, particularly 6D. The anti- SHIPGLERPS antibody also recognised chromosome 1B encoded beta-, gamma- and omega-gliadins, while the anti-METSRVPGL antibody recognised 1D encoded alpha- and beta-gliadins. The absence of sequences within the major gliadin families that are highly homologous to the latter two N-terminal LM(r)GS sequences may suggest that some monomeric LM(r)GS could exist within the electrophoretically-resolved gliadins. These antibodies will provide valuable reagents for the study of the roles of particular LM(r)GS families in the structure and function of the glutenin macropolymer, the role of different LM(r)GS types in determining the influence of allelic variation of LM(r)GS composition on dough properties, and potentially in the development of diagnostics for these flour components
Keyword	[ Hide all but 1 of 43 ] alleles allelic variation amino acid sequence antibody biotypes chinese chromosomal location chromosome cross reaction detection development dough dough properties elisa family flour gene gene location genetic variation genome gliadin glutenin glutenin subunit immunoblotting line differences loci locus low m(r) subunits m(r) macropolymer mobility monoclonal antibodies n-terminal peptides omega-gliadins peptide polypeptide repeats rye sequence substitution triticum aestivum wheat flour wheat glutenin

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