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GrainGenes Reference Report: CCM-78-79

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Reference
CCM-78-79
Title
N-terminal amino acid sequence analysis of endosperm proteins in Japanese hexaploid wheat
Journal
Cereal Chemistry
Year
2001
Volume
78
Pages
79-83
Author
Nakamura H
Abstract
Summary: Semidry electroblotting is convenient and allows a rapid and efficient protein transfer from two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) gels onto sequencer stable supports for protein microsequence analysis in a gas-phase sequencer. Using this technique, I determined the amino acid sequences of the endosperm proteins in Japanese hexaploid commercial wheats (Triticum aestivum). Based on sequence determination of the Japanese hexaploid wheats, the endosperm protein could be easily characterized. Wheat endosperm protein, extracted in the presence of 2-mercaptoethanol and SDS, fractionated into many protein polypeptides using 2D-PAGE under dissociating conditions. These components were grouped into HMW glutenin subunits, alpha-, beta- or gamma-gliadins, and novel protein polypeptides by using the N-terminal amino acid sequences. The novel endosperm protein polypeptides were detected, and two new types of N-terminal amino acid sequences have been found for protein polypeptides. These polypeptides have much faster electrophoresis mobility during 2D-PAGE and are therefore probably a much smaller size than any other peptides of endosperm protein groups found in hexaploid wheat. Ten protein polypeptides have been purified from cultivars of Japanese wheat. Some differences in the contents of amino acids for four protein polypeptide spots were apparent in Japanese wheat
Keyword
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alpha
amino acid
amino acid sequence
bread wheat
cereal
chromosome-1a
d genome
electrophoresis
endosperm
endosperm proteins
gamma
gel-electrophoresis
gels
gliadin
glutenin
glutenin subunit
hexaploid
hexaploid wheat
hmw
hmw glutenin
hmw glutenin subunit
mobility
n-terminal amino acid sequence
nucleotide
peptide
polyacrylamide
polyacrylamide-gel electrophoresis
polyacrylamide-gel-electrophoresis
polypeptide
quality
sequence
sequence analysis
subunit
supports
triticum aestivum
two-dimensional polyacrylamide gel electrophoresis
weight glutenin subunits
wheat endosperm
wheats