Publication no. C-2000-1205-02R |  VIEW ARTICLE

N-Terminal Amino Acid Sequence Analysis of Endosperm Proteins in Japanese Hexaploid Wheat.

Hiro Nakamura (1). (1) Tohoku National Agricultural Experiment Station, Morioka, Iwate 020-0198, Japan Cereal Chem. 78(1):79-83. Accepted October 5, 2000. Copyright 2001 American Association of Cereal Chemists, Inc.

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.