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GrainGenes Reference Report: JCS-30-303

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Reference	JCS-30-303
Title	Comparative studies of high M(r) subunits of rye and wheat. II. Partial amino acid sequences
Journal	Journal of Cereal Science
Year	1999
Volume	30
Pages	303-313
Author	Kipp B Wieser H
Abstract	High M(r) subunit fractions of two rye cultivars (Danko and Halo) and of the wheat cultivar Rektor were isolated from defatted flours by extraction with 50% (v/v) aqueous propan-1-ol under reducing conditions and 60 degrees C followed by precipitation using a 60% concentration of propan-1-ol, centrifugation, dialysis and lyophilisation. Two single major subunits of rye Danko were isolated by preparative RP-HPLC and characterised by N-terminal sequencing which showed a strong homology to wheat high M(r) subunits. Total high M(r) subunit fractions of Danko, Halo and Rektor were digested with TPCK-treated trypsin at 37 degrees C for 8 h. The partial hydrolysates were pre-separated into the fractions I (M(r) <10000) and II (M(r) >10000) by centrifugation on a Centriprep 10-membrane. Both fractions were then separated by reversed-phase HPLC on C18 silica gel. Dominating peptides were purified by preparative rechromatography under slightly changed HPLC conditions and characterised by amino acid analysis and determination of amino acid sequences. The resulting sequences of 19 (Danko), 12 (Halo) and 15 peptides (Rektor) were compared with known sequences from x- and y-type high M(r) subunits of wheat. With the exception of three peptides, the sequences could be assigned to the three different domains of high M(r) subunits. Even when the sequences of the rye peptides were similar to those of wheat, they were not identical in any case; on average the variations were about 20%. Most of rye peptides corresponded to the x-type subunits of wheat. Within the most characteristic sequence motifs belonging to the repetitive domain B, fundamental differences between wheat and rye were not detected
Keyword	[ Hide all but 1 of 22 ] amino acid amino acid sequence chemical composition domain extraction flour homology hplc hydrolysates kernels m(r) molecular weight peptide plant proteins rp-hplc rye rye flour secale cereale sequence triticum aestivum wheat cultivar wheat flour

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