Characterisation of the wheat Mr 15000 ?grain-softness protein? and analysis of the relationship between its accumulation in the whole seed and grain softness

Jolly, Christopher J.; Rahman, Sadequr; Kortt, A. A.; Higgins, T. J. V.

doi:10.1007/bf00838714

Cited by 133 publications

(140 citation statements)

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“…The major N-terminal sequence and sequences of peptides derived from protease digests of grain softness protein were reported by Jolly et al (1993). No similarity was found between these sequences and the N-terminal sequence found for P-30,000.…”

Section: Dtscusslonmentioning

confidence: 99%

Identification and Characterization of a Novel Arabinoxylanase from Wheat Flour

et al. 1997

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An endogenous wheat (Triticum aestivum) flour endoxylanase was purified t o homogeneity from a crude wheat flour extract by ammonium sulfate precipitation and cation-exchange chromatography. The 30-kD protein had an isoelectric point of 9.3 or higher. A sequence of 19 amino acids at the NH, terminus showed 84.2% identity with an interna1 sequence of the 15-kD grain-softness protein, friabilin. High-performance anion-exchange chromatography and gel-permeation analysis of the hydrolysis products indicated the preferential hydrolysis of highly branched structures by the enzyme; wheat arabinoxylan and rye (Secale cereale) arabinoxylan (high arabinose to xylose ratios) were hydrolyzed more efficiently by this enzyme than oat (Avena safiva) spelt xylan (low arabinose to xylose ratios). The release of the hydrolysis products as a function of time suggested that the endoxylanolytic activity was associated with the release of arabinose units from the polysaccharides, suggesting that the enzyme action i s similar to that by endoxylanases from Ceratocystis paradoxa, Aspergillus niger, and Neurospora crassa. Although the enzyme released arabinose from arabinoxylan, it did not hydrolyze pnitrophenyl-a-i-arabinofuranoside. From the above, it follows that the enzyme, called arabinoxylanase, differs from most microbial endoxylanases and from an endoxylanase purified earlier from wheat flour.AX, the main NSP in wheat (Triticum uestivum) endosperm, consists of a linear backbone of 1P-linked 6-Dxylopyranose units with L-arabinofuranosyl residues attached to the main chain by 1,3-and/or 1,2-cu-glycosidic linkages. Its structure was recently intensively studied using 'H-NMR spectroscopy and methylation analysis (Hoffmann et al., 1991(Hoffmann et al., , 1992a(Hoffmann et al., , 1992bGruppen, 1992aGruppen, , 1992bGruppen, , 1993Cleemput et al., 1993Cleemput et al., , 1995a.NSP-hydrolyzing enzymes are found in wheat grain (Preece and MacDougall, 1958;Kulp, 1968;Lee and Ronalds, 1972;Schmitz et al., 1974;Bremen, 1981; Adlung, 1985;Moore and Hoseney, 1990), in germinated wheat and wheat bran (Beldman et al., 1996), in barley (Hordeum vulgure) aleurone layers (Taiz and Honigman, 1976;Dashek

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Section: Dtscusslonmentioning

confidence: 99%

Identification and Characterization of a Novel Arabinoxylanase from Wheat Flour

et al. 1997

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“…Rosella was used as the source of GSP to raise antiserum and to obtain peptide sequences (Jolly et al, 1993). cDNA clones were isolated from libraries prepared from the hard Australian cv.…”

Section: Wheat Cultivars Usedmentioning

confidence: 99%

“…First-dimension gels were then equilibrated in 1 ml reducing buffer (SO mM Tris/HCl, pH 7.5, 3% SDS, 10% glycerol, 0.5% dithiothreitol) for 15 min, followed by 1 ml reducing buffer with 130 mM iodoacetamide for 15 min. The iodoacetamide removed an artifact due to excess reducing agent (Jolly et al, 1993). For the second dimension (SDS/ PAGE), the gels were carefully laid onto a 1.5-mm gradient SDS/PAGE gel and sealed with 1 % agarose in SDSPAGE stacking gel buffer.…”

Section: Two-dimensional Polyacrylamide Gel Electrophoresismentioning

confidence: 99%

“…For the second dimension (SDS/ PAGE), the gels were carefully laid onto a 1.5-mm gradient SDS/PAGE gel and sealed with 1 % agarose in SDSPAGE stacking gel buffer. SDS/PAGE was then performed as described (Jolly et al, 1993). A control sample of proteins extracted in SDSPAGE buffer was concurrently electrophoresed in the second dimension in a lane at the side of the gels.…”

Section: Two-dimensional Polyacrylamide Gel Electrophoresismentioning

confidence: 99%

“…Amino acid sequencing of the Nterminus of GSP purified by preparative SDSPAGE indicated that it was comprised of a major polypeptide and one or more minor polypeptides. We also reported the sequence of twelve peptides obtained from digestion of GSP by either lysyl endopeptidase or chymotrypsin (Jolly et al, 1993). Using an antiserum generated against GSP purified by SDS/ PAGE, we showed that the accumulation of GSP in Triticum aestivum seeds was dependent on the presence of the short arm of chromosome 5D.…”

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confidence: 98%

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Cloning of a wheat 15‐kDa grain softness protein (GSP)

Rahman

Jolly

Skerritt

et al. 1994

European Journal of Biochemistry

108

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The wheat starch 15-kDa protein (called grain softness protein or GSP) consists of a major polypeptide and several minor polypeptides. An antiserum raised against GSP was used to screen a wheat cDNA library. A cDNA family encoding approximately 15-kDa proteins that included a heptapeptide sequence previously isolated from protease digests of GSP was identified. A partial cDNA was used in a prokaryotic expression system to produce a fusion protein which reacted strongly against the original anti-GSP serum. A new antiserum raised against the fusion protein produced a weak reaction against a 15-kDa polypeptide extracted from wheat seeds. The results suggest that the proteins encoded by the cDNA family form a minor component of the mixture of 15-kDa polypeptides defined as GSP. RNA complementary to the cDNAs could be extracted from both soft and hard wheat grains from about half-way through grain filling. The encoded proteins are novel members of the 2s superfamily of seed proteins, a diverse family of proteins which maintain a characteristic framework of cysteine residues. The deduced proteins show the highest similarity to the oat 16-kDa avenin and to wheat puroindoline (a lipid-binding 15-kDa protein from wheat). Review of previously published data shows that puroindoline is also closely related to the major polypeptide of GSP, suggesting that the lipid-binding properties of GSP polypeptides may influence grain softness.

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Starch Granule-Associated Proteins and Polypeptides: A Review

2001

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This paper reviews current information regarding the (non-gluten) proteins which are naturally located in and on starch granules, and which are termed starch granule associated proteins (SGAPs). At least ten major SGAPs can be extracted from most starches and have molecular weights in the range of approximately 5 to 149 kDa. A substantial number of these proteins are located at the starch granule surface, where their presence in association with that of other minor granule components (such as lipids), appears to significantly influence the overall properties of both starch granules and starchy products. The different extraction conditions which can be exploited to selectively remove SGAPs are detailed. Although some SGAPs may serve as supplementary storage proteins, the majority of SGAPs are believed to be starch biosynthetic enzymes, a proportion of which remain trapped within the growing granule structure. The identity and quantities of the biosynthetic enzymes varies with botanical source, genetic variety and even with time. Current knowledge regarding the identities, compositions, locations, properties and functions of several of the more common SGAPs (e.g. the ~15 kDa friabilin/puroindoline proteins, the ~30 kDa protein, and the ~60 kDa starch granule-bound starch synthase (SGBSS) protein) are reviewed in detail.

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Characterisation of the wheat Mr 15000 ?grain-softness protein? and analysis of the relationship between its accumulation in the whole seed and grain softness

Cited by 133 publications

References 21 publications

Identification and Characterization of a Novel Arabinoxylanase from Wheat Flour

Identification and Characterization of a Novel Arabinoxylanase from Wheat Flour

Cloning of a wheat 15‐kDa grain softness protein (GSP)

Starch Granule-Associated Proteins and Polypeptides: A Review

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