Complete amino acid sequence of puroindoline, a new basic and cystine‐rich protein with a unique tryptophan‐rich domain, isolated from wheat endosperm by Triton X‐114 phase partitioning

Blochet, J.E.; Chevalier, Catherine; Forest, Éric; Pebay‐Peyroula, Eva; Gautier, Marie‐Françoise; Joudrier, Philippe; Pézolet, Michel; Marion, Didier

doi:10.1016/0014-5793(93)80249-t

Cited by 207 publications

(175 citation statements)

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“…Puroindolines were purified from wheat seeds according to the method described previously by Blochet et al (1993) and by Wilde et al (1993). In addition, in order to remove salt contamination from puroindolines, solutions were centrifuged on an Amicon 3000 filter at 7000g for 1 h. Centrifugations were first done with 150 mM NaCl and then with pure distilled water.…”

Section: Methodsmentioning

confidence: 99%

“…Puroindolines are water-soluble basic and cysteine-rich proteins of about 13 kDa that have been isolated from wheat endosperm using Triton X-114 phase partitioning (Blochet et al, 1991(Blochet et al, , 1993. Two isoforms named puroindoline-a and puroindoline-b have been purified and characterized.…”

mentioning

confidence: 99%

“…Two isoforms named puroindoline-a and puroindoline-b have been purified and characterized. They exhibit more than 50% homology in their amino acid sequence (Blochet et al, 1991;Gautier et al, 1994). Puroindoline-a, the first characterized isoform, contains a unique tryptophan-rich domain (WRWWKWWK) which is at the origin of the name of this protein (from the greek puros for wheat and indoline for the indole ring of tryptophan).…”

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confidence: 99%

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Determination of the Secondary Structure and Conformation of Puroindolines by Infrared and Raman Spectroscopy

Bihan

Désormeaux

et al. 1996

Biochemistry

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The conformation of puroindoline-a and -b, two basic lipid-binding proteins isolated from wheat seedlings, has been studied for the first time by infrared and Raman spectroscopy. The infrared results show that puroindoline-a and -b have similar secondary structure composed of approximately 30% R-helices, 30% -sheets, and 40% unordered structure at pH 7. The conformation of both puroindolines is significantly pH-dependent. The reduction of the disulfide bridges leads to a decrease of the solubility of puroindolines in water and to an increase of the -sheet content by about 15% at the expense of the R-helix content. Raman spectroscopy confirms the structure similarity between the two puroindolines with little differences in the side chains' environment. All the disulfide bridges are in a gauche-gauchegauche conformation, and the unique tyrosine residue present in both puroindolines is hydrogen-bonded to water. Raman spectra have been recorded in both H 2 O and D 2 O media, thus providing additional information concerning the accessibility of certain residues to water. We have also observed that puroindoline-a tends to form some aggregates under acidic and high ionic strength conditions. Nearultraviolet circular dichroism measurements suggest that the tryptophan-rich domain is involved in this aggregate formation. Finally, on the basis of a combined infrared and sequence conformational analysis, we propose a secondary structure assignment for both puroindolines. The results show that puroindolines exhibit a similar folding pattern with plant nonspecific lipid-transfer protein and some amylase-protease inhibitors. These proteins could form a homogeneous structural family of plant proteins involved in the defense against pathogens that are probably derived from a common "helicoidal" protein ancestor.

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Section: Methodsmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Determination of the Secondary Structure and Conformation of Puroindolines by Infrared and Raman Spectroscopy

Bihan

Désormeaux

et al. 1996

Biochemistry

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“…In all cases these procedures led to the isolation of low molecular and cysteine-rich proteins including thionins and amylase inhibitors [14][15]. Since 1990, using mild biochemical extraction and fractionation procedures, two main lipid binding proteins families were isolated from wheat flour, lipid transfer proteins and puroindolines [16][17]. Both protein families belong to albumins.…”

Section: From Lipid-protein Interactions To Lipid-binding Proteinsmentioning

confidence: 99%

Functionality of lipids and lipid-protein interactions in cereal-derived food products

Marion

Dubreil

Douliez

2003

OCL

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Résumé :Les lipides et en particulier les lipides des céréales jouent un rôle important dans la transformation et la qualité des produits céréaliers de cuisson (pains, biscuits) et des boissons (bière). La plupart des mécanismes physico-chimiques responsables de la fonctionnalité des lipides a été étudiée et récemment, le rôle particulier des protéines impliquées dans la fixation des lipides, les protéines de transfert de lipides et les puroindolines a été mis en évidence. L'état des recherches menées dans ce domaine est présenté dans cette revue et les informations acquises aujourd'hui montrent que de nouvelles perspectives sont envisageables pour la sélection et la transformation en vue d'améliorer la qualité des céréales et des produits céréaliers.Summary : Lipids and especially cereal lipids play a significant role in the processing and quality of cereals and baked cereal foods (bread, biscuits) and beverages (beer). Most of the physico-chemical mechanisms responsible for the lipid functionality has been investigated and recently the specific role of lipid-binding proteins, e.g. lipid transfer proteins and puroindolines, has been highlighted. The state of the researches performed in this field are briefly presented in this review and the data obtained until now show that new perspectives are opened in cereal breeding and processing for improving the quality of cereals and cereal products.

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“…Wheat friabilin proteins were extracted using the Triton X-114 (TX-114) phase partitioning method of Blochet et al (1993). The extracts were stored at 4°C.…”

Section: Friabilin Extractionmentioning

confidence: 99%

In Vitro Starch Binding Experiments: Study of the Proteins Related to Grain Hardness of Wheat

Bakó

Gárdonyi

Tamás

Developments in Plant Breeding

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SummaryTwo friabilin components, puroindoline a and GSP-1 were expressed in Escherichia coli. Starch binding properties of the recombinant polypeptides and of friabilin extracted from wheat flour were compared in vitro. The produced proteins as well as native wheat friabilin bound to starch granules prepared from different (soft, hard and durum) wheat cultivars. Starch granules also bound specifically several wheat endosperm proteins other than friabilin. IntroductionGrain hardness (endosperm texture) forms the fundamental basis of commercial differentiation of wheat cultivars. It is possible to differentiate 'soft' and 'hard' hexaploid wheats and 'very hard' durum wheat as three distinct qualitative classes. Texture determines flour particle size, starch damage, water absorption and milling yield. Therefore, grain hardness is an indicator of the suitability of a particular flour for a particular product. To the grower, texture is important as generally higher premiums are paid for the harder wheats (Morris, 2002).The hardness of the grain is largely determined by the properties of the endosperm. The surface of the starch granules is covered by lipids and special proteins. Hard wheat starches prepared by water sedimentation have more material adhering to their surface than soft wheat starches prepared by the same method when examined by scanning electron microscopy (Barlow et al., 1973). It is generally accepted that the adhesion between the granules and the protein matrix is stronger in hard wheats than in soft wheats. During the milling of hard wheat, starch granules fragmentize. This fragmentation is called starch damage. Starch damage is the most important factor in determining water absorption of flour. It also affects the amount of carbohydrates available to yeasts, the fermentative activity, gas production, loaf volume and, as a result, baking quality. So far the most well characterized source of variation in grain hardness is the Ha (Hardness) locus located on the short arm of chromosome 5D of hexaploid wheat. The genes for puroindoline a, puroindoline b and GSP-1 (the three major components of the friabilin protein fraction) are tightly linked to the Ha locus. There is an unbroken linkage between mutations in any of the puroindoline genes and grain hardness. Friabilin is abundant on the surface of water-washed soft wheat starch granules, scarce on hard wheat starch and absent on durum wheat starch.The biochemical mechanism governing endosperm texture is poorly understood. The cause of the strong adhesion between starch granules and protein matrix in hard and durum wheat cultivars is unknown. How friabilin located on the surface of the granules impairs the adhesion in soft wheats is also unclear. Detailed knowledge on the molecular background of grain hardness could help in producing cultivars specially fit for any one particular purpose.Isolation of the components located at the starch-gluten interface and investigation of their interactions in in vitro experiments might help to elucidate the mechanism ...

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Complete amino acid sequence of puroindoline, a new basic and cystine‐rich protein with a unique tryptophan‐rich domain, isolated from wheat endosperm by Triton X‐114 phase partitioning

Cited by 207 publications

References 27 publications

Determination of the Secondary Structure and Conformation of Puroindolines by Infrared and Raman Spectroscopy

Determination of the Secondary Structure and Conformation of Puroindolines by Infrared and Raman Spectroscopy

Functionality of lipids and lipid-protein interactions in cereal-derived food products

In Vitro Starch Binding Experiments: Study of the Proteins Related to Grain Hardness of Wheat

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