scholarly journals Conformational differences between two wheat (Triticum aestivum) ‘high-molecular-weight’ glutenin subunits are due to a short region containing six amino acid differences

1989 ◽  
Vol 263 (3) ◽  
pp. 837-842 ◽  
Author(s):  
A P Goldsbrough ◽  
N J Bulleid ◽  
R B Freedman ◽  
R B Flavell
Keyword(s):  
Molecular Weight ◽  
Triticum Aestivum ◽  
Amino Acid ◽  
High Molecular Weight ◽  
Polyacrylamide Gel Electrophoresis ◽  
Amino Acid Sequences ◽  
Glutenin Subunits ◽  
Sds Page ◽  
Hmw Glutenin

‘High-molecular-weight’ (HMW, high-Mr) glutenin subunits are protein constituents of wheat (Triticum aestivum) seeds and are responsible in part for the viscoelasticity of the dough used to make bread. Two subunits, numbered 10 and 12, are the products of allelic genes. Their amino acid sequences have been derived from the nucleic acid sequences of the respective genes. Subunit 10 has fewer amino acids than subunit 12, but migrates more slowly on SDS/PAGE (polyacrylamide-gel electrophoresis). This anomaly is due to between one and six of the amino acid differences between the subunits, localized towards the C-terminal end of the proteins. This has been established by making chimaeric genes between the genes for subunits 10 and 12, transcribing and translating them in vitro and analysing the products by SDS/PAGE. The postulated conformational differences between subunits 10 and 12 are discussed in relation to current hypotheses for the structure of HMW glutenin subunits.

An approach for isolating high-molecular-weight glutenin subunit genes using monoclonal antibodies

Genome ◽  
2006 ◽  
Vol 49 (2) ◽  
pp. 181-189 ◽  
Author(s):  
H Q Wang ◽  
X Y Zhang
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibody ◽  
Triticum Aestivum ◽  
Amino Acid ◽  
High Molecular Weight ◽  
Storage Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Glutenin Subunits ◽  
Sds Page

High-molecular-weight glutenin subunits (HMW-GSs) play an important role in the breadmaking quality of wheat flour. In China, cultivars such as Triticum aestivum 'Xiaoyan No. 6' carrying the 1Bx14 and 1By15 glutenin subunits usually have attributes that result in high-quality bread and noodles. HMW-GS 1Bx14 and 1By15 were isolated by preparative sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and used as an antigen to immunize BALB/c mice. A resulting monoclonal antibody belonging to the IgG1 subclass was shown to bind to all HMW-GSs of Triticum aestivum cultivars, but did not bind to other storage proteins of wheat seeds in a Western blot analysis. After screening a complementary DNA expression library from immature seeds of 'Xiaoyan No. 6' using the monoclonal antibody, the HMW-GS 1By15 gene was isolated and fully sequenced. The deduced amino acid sequence showed an extra stretch of 15 amino acid repeats consisting of a hexapeptide and a nonapeptide in the repetitive domain of this y-type HMW subunit. Bacterial expression of a modified 1By15 gene, in which the coding sequence for the signal peptide was removed and a BamHI site eliminated, gave rise to a protein with mobility identical to that of HMW-GSs extracted from seeds of 'Xiaoyan No. 6' via SDS-PAGE. This approach for isolating genes using specific monoclonal antibody against HMW-GS genes is a good alternative to the extensively used polymerase chain reaction (PCR) technology based on sequence homology of HMW-GSs in wheat and its relatives.Key words: wheat, HMW-GS, monoclonal antibody, immunoscreen.

Genetic and biochemical characterization of novel low molecular weight glutenin subunits in wheat (Triticum aestivum L.)

Genome ◽  
1997 ◽  
Vol 40 (1) ◽  
pp. 41-48 ◽  
Author(s):  
Guttapadu Sreeramulu ◽  
Nagendra K. Singh
Keyword(s):  
Molecular Weight ◽  
Triticum Aestivum ◽  
Amino Acid ◽  
Biochemical Characterization ◽  
Amino Acid Sequences ◽  
Gene Location ◽  
Relative Molecular Mass ◽  
Low Molecular Weight ◽  
Glutenin Subunits ◽  
Moving Band

Two novel low molecular weight subunits of glutenin with relative molecular mass (Mr values) of 30 and 32 kDa were isolated from the seeds of hexaploid wheat and characterized at genetic and biochemical levels. Among 115 Indian bread wheat cultivars analysed, 40 had a narrow doublet of the new protein bands, 69 had a wide doublet, 3 had only the faster moving band of the doublet, and the remaining 3 cultivars had only the slower moving band. These subunits could be seen in the alkylated glutenin preparations only and the genes for the faster (designated Glu-D4) and slower (designated Glu-D5) moving protein bands of the doublet were located on chromosomes 1D and 7D, respectively. Amino acid composition of the two new subunits was quite different from those of the other well-characterized gluten proteins, as the new subunits have lower amounts of proline and relatively higher amounts of glycine, aspartic acid – asparagine, cysteine, and lysine. Polyclonal antibodies raised against these polypeptides cross-reacted strongly with the major low molecular weight subunits of wheat glutenin (Glu-3 subunits), but did not cross-react with the high molecular weight glutenin subunits or monomeric gliadins. Furthermore, preliminary results on the N-terminal amino acid sequences of the new subunits show homology with the major low molecular weight glutenin subunits, suggesting an evolutionary link between the two.Key words: Triticum aestivum, glutenin subunits, gene location, immunoblotting.

scholarly journals Identification of Glu-A1 and Glu-D1 High Molecular Weight Glutenin Subunits of Common Wheat (Triticum aestivum L.) Using Genetic Markers

2018 ◽  
Vol 72 (6) ◽  
pp. 349-357
Author(s):  
Marina Tikhonova ◽  
Anne Ingver ◽  
Reine Koppel
Keyword(s):  
Molecular Weight ◽  
Triticum Aestivum ◽  
High Molecular Weight ◽  
Common Wheat ◽  
Triticum Aestivum L ◽  
Glutenin Subunits ◽  
Bread Making ◽  
Accurate Identification ◽  
Sds Page ◽  
Pcr Method

Abstract High molecular weight glutenin subunits (HMW-GS) of wheat are important factors in the determination of bread-making quality. They are responsible for elasticity and polymer formation of wheat dough. In the present study, 43 winter and 40 spring common wheat (Triticum aestivum L.) cultivars originated from Estonia, Belarus, Finland, Denmark, France, Germany, the Great Britain, Latvia, Lithuania, the Netherlands, Norway, Poland, Russia, Sweden, and New Zealand were characterised for Glu-A1 and Glu-D1 allelic composition using PCR method. Analyses were conducted with one DNA marker for identification of Glu-D1 allele encoding subunit Dx5, three DNA markers for Glu-A1 Ax1, Ax2* and AxNull subunits. It was determined that 32 (74.4%) winter and 35 (83.3%) spring cultivars had allele Glu-D1d, and 23 (53.5%) winter and 33 (78.6%) spring — Glu-A1a or Glu-A1b alleles, which have positive effect on dough properties. Polymorphism at Glu-A1 locus was detected in 15 cultivars, and 9 cultivars were polymorphic for locus Glu-D1. The obtained results were compared with published SDS-PAGE data. Complete or partial agreements were found for 78.1% of Glu-A1 and 70.6% of Glu-D1 alleles. Rapid and accurate identification of wheat Glu-1 alleles by molecular markers can be used for selection of wheat genotypes with good bread-making potential.

Analysis of high-molecular-weight glutenin subunits in five amphidiploids and their parental diploid species Aegilops umbellulata and Aegilops uniaristata

2014 ◽  
Vol 13 (2) ◽  
pp. 186-189 ◽  
Author(s):  
Shoufen Dai ◽  
Li Zhao ◽  
Xiaofei Xue ◽  
Yanni Jia ◽  
Dengcai Liu ◽  
...  
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Triticum Turgidum ◽  
Chromosome Doubling ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Diploid Species ◽  
Glutenin Subunits ◽  
Aegilops Umbellulata ◽  
Sds Page

Amphidiploids serve as a bridge for transferring genes from wild species into wheat. In this study, five amphidiploids with AABBUU and AABBNN genomes were produced by spontaneous chromosome doubling of unreduced triploid F1 gametes from crosses between diploid Aegilops (A. umbellulata accessions CIae 29 and PI 226500, and A. uniaristata accession PI 554419) and tetraploid Triticum turgidum (ssp. durum cultivar Langdon and ssp. dicoccum accessions PI 94 668 and PI 349045) species. The composition of high-molecular-weight glutenin subunits (HMW-GS) in these amphidiploids and in their parental A. umbellulata and A. uniaristata species was analysed. As expected, the amphidiploids from T. turgidum ssp. dicoccum accession PI 944668 or PI 349045 and A. umbellulata accession CIae 29 or PI 226500 and A. uniaristata accession PI 554419 showed the same HMW-GS patterns as those of their Aegilops parents, because HMW-GS genes were all silenced in the T. turgidum ssp. dicoccum parents. The amphidiploids from CIae 29 and Langdon inherited all of the HMW-GS genes from their parents except for the Uy type. Using 10 and 15% sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and 10% urea/SDS–PAGE, 11 Ux and ten Uy types in 16 combinations were observed in 48 A. umbellulata accessions, and two Nx and two Ny types in two combinations were detected in six A. uniaristata accessions. These novel HMW-GS variants may provide new genetic resources for improving the quality of wheat.

scholarly journals Composition of High Molecular Weight Glutenin Subunits in Polish Common Wheat Cultivars (Triticum aestivum L.)

2018 ◽  
Vol 2018 ◽  
pp. 1-8 ◽  
Author(s):  
Ewa Filip
Keyword(s):  
Molecular Weight ◽  
Triticum Aestivum ◽  
High Molecular Weight ◽  
Wheat Gluten ◽  
Triticum Aestivum L ◽  
Glutenin Subunits ◽  
Bread Making ◽  
New Information ◽  
Sds Page ◽  
Genes Encoding

The main goal of our study was to present research data on genes encoding high molecular weight glutenin subunits (HMW-GS) associated with high flour bread-making quality. This is the leading research objective in our institute in the area of wheat gluten in cultivars that have not been studied so far in that respect, but which can potentially be a valuable source of new information. Identification and characterization of high molecular weight glutenin subunits (HMW-GS) were performed using sequencing and SDS-PAGE and STS-PCR methods. Genes located in the vicinity of the Glu-1 locus have been identified and characterized in 28 Polish cultivars of Triticum aestivum. The results were then analyzed using the following computer programs: Finch TV, BLAST, MEGA 4, Molecular Imager® Gel Doc™ XR, and Quantity One software (Bio-Rad). Three alleles (a, b, c) have been identified in the Glu-A1 locus, 6 alleles (a, b, c, d, e, k) in the Glu-B1 locus, and 2 alleles (a, d) in Glu-D1 using the SDS-PAGE method. The amplification of specific HMW-GS sequences generated one product of 450 bp in 1Dx5 in 13 cultivars of old wheat and of 435 bp in 1Dx2 in 15 cultivars. The amplification products of primers for 1Dy10 and 1Dy12 genes were 422 bp and 552 bp in size, respectively.

scholarly journals Correlation between high molecular weight gluten subunits composition and bread-making quality in Brazilian wheat

1997 ◽  
Vol 20 (4) ◽  
pp. 667-671 ◽  
Author(s):  
Ivan Schuster ◽  
Moacil Alves de Souza ◽  
Antônio Américo Cardoso ◽  
Carlos Sigueyuki Sediyama ◽  
Maurílio Alves Moreira
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Sodium Dodecyl ◽  
Glutenin Subunits ◽  
Bread Making ◽  
Hmw Glutenin Subunits ◽  
Statistical Correlations ◽  
Sds Page ◽  
Half Seed ◽  
Hmw Glutenin

Bread-making quality is one of the most important targets in the genetic improvement of wheat. Although extensive analyses of quality traits such as farinography, sodium dodecyl sulfate (SDS) sedimentation, alveography, and baking are made in breeding programs, these analyses require high amounts of seeds which are obtained only in late generations. In this experiment the statistical correlations between the high molecular weight subunit of glutenin and bread-making quality measured by alveograph, farinograph and SDS sedimentation were evaluated. Seventeen wheat genotypes were grown under the same conditions, each producing about 1 kg of seeds for the evaluations. The high molecular weight (HMW) glutenin subunits were analyzed by SDS-PAGE. Statistical correlations were highly significant between HMW glutenin subunits and alveograph and SDS sedimentation. These results indicate the possibility of manipulating major genes for wheat seed quality by coupling traditional breeding with non-destructive single seed analysis. Only half seed is necessary to perform the SDS-PAGE analysis. Therefore, the other half seed can be planted to generate the progeny. Seed yield and SDS sedimentation were statistically correlated, indicating the possibility of simultaneous selection for both traits

scholarly journals Isolation and Characterization of Two Proteins fromMoraxella catarrhalis That Bear a Common Epitope

1998 ◽  
Vol 66 (9) ◽  
pp. 4374-4381 ◽  
Author(s):  
John C. McMichael ◽  
Michael J. Fiske ◽  
Ross A. Fredenburg ◽  
Deb N. Chakravarti ◽  
Karl R. VanDerMeid ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Bacterial Attachment ◽  
Vaccine Candidates ◽  
Isolation And Characterization ◽  
Sds Page

ABSTRACT The UspA1 and UspA2 proteins of Moraxella catarrhalisare potential vaccine candidates for preventing disease caused by this organism. We have characterized both proteins and evaluated their vaccine potential using both in vitro and in vivo assays. Both proteins were purified from the O35E isolate by Triton X-100 extraction, followed by ion-exchange and hydroxyapatite chromatography. Analysis of the sequences of internal peptides, prepared by enzymatic and chemical cleavage of the proteins, revealed that UspA1 and UspA2 exhibited distinct structural differences but shared a common sequence including an epitope recognized by the monoclonal antibody 17C7. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), purified UspA1 exhibited a molecular weight of approximately 350,000 when unheated and a molecular weight of 100,000 after being heated for 10 min at 100°C. In contrast, purified UspA2 exhibited an apparent molecular weight of 240,000 by SDS-PAGE that did not change with the length of time of heating. Their sizes as determined by gel filtration were 1,150,000 and 830,000 for UspA1 and UspA2, respectively. Preliminary results indicate the proteins have separate functions in bacterial pathogenesis. Purified UspA1 was found to bind HEp-2 cells, and sera against UspA1, but not against UspA2, blocked binding of the O35E isolate to the HEp-2 cells. UspA1 also bound fibronectin and appears to have a role in bacterial attachment. Purified UspA2, however, did not bind fibronectin but had an affinity for vitronectin. Both proteins elicited bactericidal antibodies in mice to homologous and heterologous disease isolates. Finally, mice immunized with each of the proteins, followed by pulmonary challenge with either the homologous or a heterologous isolate, cleared the bacteria more rapidly than mock-immunized mice. These results suggest that UspA1 and UspA2 serve different virulence functions and that both are promising vaccine candidates.

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