The purification and N-terminal amino acid sequence analysis of the high molecular weight gluten polypeptides of wheat

Shewry, Peter R.; Field, Jacqueline; Faulks, Audrey J.; Parmar, S.; Miflin, B. J.; Dietler, Mary D.; Lew, Ellen J-L.; Kasarda, Donald D.

doi:10.1016/0167-4838(84)90293-0

Cited by 84 publications

(38 citation statements)

References 34 publications

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“…Thus, a number of different subunits can be recognised which are numbered in accordance with their mobility in SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis). 6 Although the N-terminal amino acid sequences of a number of HMW subunits were initially reported, 7 the determination of complete sequences awaited the isolation of genes. As a result the sequences of a number of proteins are now available including 1Ax, 1Bx, 1Dx, 1By and 1Dy subunits.…”

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confidence: 99%

Structural studies of glutenin subunits 1Dy10 and 1Dy12 by matrix‐assisted laser desorption/ionisation mass spectrometry and high‐performance liquid chromatography/electrospray ionisation mass spectrometry

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Foti

Saletti

et al. 2003

Rapid Comm Mass Spectrometry

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Structural studies of the high molecular weight (HMW) glutenin subunits 1Dy10 and 1Dy12 of bread wheat were conducted using matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOFMS) and reversed-phase high-performance liquid chromatography/electrospray ionisation mass spectrometry (RP-HPLC/ESI-MS). For both proteins, MALDI-TOFMS analysis showed that the isolated fractions contained a second component with a mass about 500-540 Da lower than the major component. The testing and correction of the gene-derived amino acid sequences of both proteins were performed by direct MALDI-TOFMS analysis of their tryptic peptide mixture and analysis of the digests was performed by recording several MALDI mass spectra of the mixture at low, medium and high mass ranges, optimising the matrix and the acquisition parameters for each mass range. Complementary data were obtained by RP-HPLC/ESI-MS analysis of the tryptic digest. This resulted in the coverage of the whole protein sequences except for two short fragments (T1 and T8), which are identical in the two homologous subunits, and for an additional dipeptide (T14) in subunit 1Dy12, which were not detected. It also demonstrated that, in contrast to the gene-derived data, the sequence of subunit 1Dy12 does not include the dipeptide Gly-Gln between residues Gln(454) and Pro(455), and that the lower mass components present in both fractions correspond to the same sequences lacking short peptides that are probably lost from the protein N- or C-termini. Finally, the results obtained provide evidence for the lack of a substantial level of glycosylation or other post-translational modifications of the two subunits, and demonstrate that mass spectrometric mapping is the most useful method presently available for the direct verification of the gene-derived sequences of HMW glutenin subunits and similar proteins.

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confidence: 99%

Structural studies of glutenin subunits 1Dy10 and 1Dy12 by matrix‐assisted laser desorption/ionisation mass spectrometry and high‐performance liquid chromatography/electrospray ionisation mass spectrometry

Cunsolo

Foti

Saletti

et al. 2003

Rapid Comm Mass Spectrometry

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“…As most of wheat endosperm proteins, glutenins are rich in glutamine and proline, but unlike gliadins, contain up to 20% glycine. N-terminal amino acid sequences of several glutenin subunits have been determined (3). Wheat developing endosperm poly(A)+ RNA was shown to contain mRNAs coding for glutenins (4).…”

Section: Introductionmentioning

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A wheat HMW glutenin subunit gene reveals a highly repeated structure

et al. 1985

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“…3 The 58-kDa peptide is a peptide with a relative molecular mass (M r ) of 58 kDa, and it corresponds to most of the central repetitive domain of 1Dx5 from residue 103 to residue 643. It was expressed from a gene construct in Escherichia coli; for expression purposes, the residue corresponding to serine position 102 of 1Dx5 was altered to become methionine 1 of the 58-kDa peptide.…”

Section: Methodsmentioning

confidence: 99%

“…The amino acid compositions of HMW glutenins are generally characterized by high contents of glutamine (ϳ 35 mol %), glycine (ϳ 20 mol %), and proline (ϳ 10 mol %). 3,4 It has been suggested that the type of secondary structure adopted by HMW subunits upon hydration and the network formed may both contribute to the mechanism of dough viscoelasticity. This network reportedly involves relatively ordered regions of intermolecular hydrogen bonding coexisting with disordered unbound regions.…”

Section: Introductionmentioning

confidence: 99%

Study of wheat high molecular weight 1Dx5 subunit by ¹³C and ¹H solid‐state NMR. II. Roles of nonrepetitive terminal domains and length of repetitive domain

et al. 2002

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This work follows a previous article that addressed the role of disulfide bonds in the behavior of the 1Dx5 subunit upon hydration. Here the roles of nonrepetitive terminal domains present and the length of the central repetitive domain in the hydration of 1Dx5 are investigated. This was achieved by comparing the hydration behavior of suitable model samples determined by (13)C- and (1)H-NMR: an alkylated 1Dx5 subunit (alk1Dx5), a recombinant 58-kDa peptide corresponding to the central repetitive domain of 1Dx5 (i.e., lacking the terminal domains), and two synthetic peptides (with 6 and 21 amino acid residues) based on the consensus repeat motifs of the central domain. The (13)C cross-polarization and magic angle spinning (MAS) experiments recorded as a function of hydration gave information about the protein or peptide fractions resisting plasticization. Conversely, (13)C single pulse excitation and (1)H-MAS gave information on the more plasticized segments. The results are consistent with the previous proposal of a hydrated network held by hydrogen-bonded glutamines and possibly hydrophobic interactions. The nonrepetitive terminal domains were found to induce water insolubility and a generally higher network hindrance. Shorter chain lengths were shown to increase plasticization and water solubility. However, at low water contents, the 21-mer peptide was characterized by higher hindrance in the megahertz and kilohertz frequency ranges compared to the longer peptide; and a tendency for a few hydrogen-bonded glutamines and hydrophobic residues to remain relatively hindered was still observed, as for the protein and large peptide. It is suggested that this ability is strongly dependent on the peptide primary structure.

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The purification and N-terminal amino acid sequence analysis of the high molecular weight gluten polypeptides of wheat

Cited by 84 publications

References 34 publications

Structural studies of glutenin subunits 1Dy10 and 1Dy12 by matrix‐assisted laser desorption/ionisation mass spectrometry and high‐performance liquid chromatography/electrospray ionisation mass spectrometry

Structural studies of glutenin subunits 1Dy10 and 1Dy12 by matrix‐assisted laser desorption/ionisation mass spectrometry and high‐performance liquid chromatography/electrospray ionisation mass spectrometry

A wheat HMW glutenin subunit gene reveals a highly repeated structure

Study of wheat high molecular weight 1Dx5 subunit by ¹³C and ¹H solid‐state NMR. II. Roles of nonrepetitive terminal domains and length of repetitive domain

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The purification and N-terminal amino acid sequence analysis of the high molecular weight gluten polypeptides of wheat

Cited by 84 publications

References 34 publications

Structural studies of glutenin subunits 1Dy10 and 1Dy12 by matrix‐assisted laser desorption/ionisation mass spectrometry and high‐performance liquid chromatography/electrospray ionisation mass spectrometry

Structural studies of glutenin subunits 1Dy10 and 1Dy12 by matrix‐assisted laser desorption/ionisation mass spectrometry and high‐performance liquid chromatography/electrospray ionisation mass spectrometry

A wheat HMW glutenin subunit gene reveals a highly repeated structure

Study of wheat high molecular weight 1Dx5 subunit by 13C and 1H solid‐state NMR. II. Roles of nonrepetitive terminal domains and length of repetitive domain

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Study of wheat high molecular weight 1Dx5 subunit by ¹³C and ¹H solid‐state NMR. II. Roles of nonrepetitive terminal domains and length of repetitive domain