The Biosynthesis, Structure and Properties of Napin - the Storage Protein From Rape Seeds

Byczyńska, Agnieszka; Barciszewski, Jan

doi:10.1016/s0176-1617(99)80277-6

Cited by 21 publications

(18 citation statements)

References 50 publications

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“…They are encoded by a multigenic family, initially synthetised as a precursor [16] which is proteolytically cleaved to generate mature napin chains. Only four napin genes were sequenced [17][18][19][20][21][22] among 10-20 napin genes attended. Such a large number of genes result in a multiplicity of isoforms among those five were identified by Monsalve and Rodriguez [11].…”

Section: Introductionmentioning

confidence: 99%

Large scale purification of rapeseed proteins (Brassica napus L.)

Bérot

Compoint

Larré

et al. 2005

Journal of Chromatography B

135

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Section: Introductionmentioning

confidence: 99%

Large scale purification of rapeseed proteins (Brassica napus L.)

Bérot

Compoint

Larré

et al. 2005

Journal of Chromatography B

135

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“…The economic interest on Brassicaceae seeds has increased in the last decade since Brassica represents one of the most important oil seed annual crops in the world, as well as one of the main sources for animal nutrition. Their 2S albumins represent a good model for studying expression and maturation processes in plant tissues [1]. The 2S albumin class is an abundant group of seed storage proteins widely distributed in numerous species, which have been isolated and characterized from several Brassicaceae as Sinapis alba (yellow mustard), Brassica juncea (oriental mustard), Raphanus sativus (radish), Ricinus communis (castor bean), Arabidopsis thaliana (thale cress) and Brassica napus (rapeseed) [2][3][4][5][6][7].…”

mentioning

confidence: 99%

Recombinant pronapin precursor produced in Pichia pastoris displays structural and immunologic equivalent properties to its mature product isolated from rapeseed

Palomares¹,

Monsalve²,

Rodrı́guez³

et al. 2002

European Journal of Biochemistry

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2S albumin storage proteins from rapeseed (Brassica napus), called napins, consist of two different polypeptide chains linked by disulphide bridges, which are derived by proteolytic cleavage from a single precursor. The precursor form of the napin BnIb (proBnIb) has been cloned using a PCR strategy and sequenced. The amino-acid sequence deduced from the clone includes 31 residues of the small chain and 75 of the large chain, which are connected by the peptide SerGlu-Asn. Expression of the cDNA encoding proBnIb has been carried out in the methylotrophic yeast Pichia pastoris. The induced protein was secreted to the extracellular medium at a yield of 80 mgAEL )1 of culture and was purified by means of size-exclusion chromatography and reverse phase-HPLC. Recombinant proBnIb appeared properly folded as its molecular and spectroscopic properties were equivalent to those of the mature heterodimeric protein.As 2S albumin storage proteins from Brassicaceae have been shown to be type I allergy inducers, the immunological activity of the recombinant proBnIb was analysed as a measure of its structural integrity. The immunological properties of the recombinant precursor and the natural napin were indistinguishable by immunoblotting and ELI-SA inhibition using polyclonal antisera and sera of patients allergic to mustard and rapeseed. In conclusion, the recombinant expression of napin precursors in P. pastoris has been shown to be a successful method for high yield production of homogeneous and properly folded proteins whose polymorphism and complex maturation process limited hitherto their availability.Keywords: 2S albumin; rapeseed; allergen; Pichia pastoris; BnIb precursor.The economic interest on Brassicaceae seeds has increased in the last decade since Brassica represents one of the most important oil seed annual crops in the world, as well as one of the main sources for animal nutrition. Their 2S albumins represent a good model for studying expression and maturation processes in plant tissues [1]. The 2S albumin class is an abundant group of seed storage proteins widely distributed in numerous species, which have been isolated and characterized from several Brassicaceae as Sinapis alba (yellow mustard), Brassica juncea (oriental mustard), Raphanus sativus (radish), Ricinus communis (castor bean), Arabidopsis thaliana (thale cress) and Brassica napus (rapeseed) [2][3][4][5][6][7]. The 2S albumins from B. napus, called napins, are encoded by multigene families, whose products exhibit a high degree of sequence similarity. Members of this family constitute small (12-15 kDa) and basic (around pI 11.0) proteins composed of two different chains (small and large) linked by disulphide bridges, which are expressed as a single polypeptide precursor (preproprotein) [8]. The internal processed fragment (IPF), which connects both chains in the precursor, is eliminated by proteolytical cleavage together with N-and C-terminal extensions during the post-translational maturation of the preproprotein [1,[8][9][10]. The most common and ab...

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“…Members of this family constitute small, 12 to15 kDa, and basic proteins composed of two different chains (small and large) linked by disulphide bridges and are expressed as precursors/ preproteins (Crouch et al 1983). They undergo proteolytic cleavages at one or more sites during maturation (Byczynska and Barciszewski 1999). Several functions or activities have been assigned to this family of proteins: nitrogen and sulphur storage (Higgins 1984), antifungal activity (Terras et al 1993), calmodulin antagonist activity (Polya et al 1993), allergenicity (Monsalve et al 1993).…”

Section: Resultsmentioning

confidence: 99%

Extracellular localization of napin in the embryogenic tissues of Brassica napus spp. oleifera

Namasivayam

Skepper

Hanke

2008

In Vitro Cell.Dev.Biol.-Plant

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Napin, a storage protein, has been reported to be transcribed abundantly during the pre-embryogenic stage and associated with the induction of Brassica napus secondary embryogenesis. In this study, we studied the distribution pattern of napin in the winter oilseed rape embryogenic tissue in comparison to that of the nonembryogenic tissue using the indirect immunofluorescence localisation coupled with the ultrastructural immunogold labelling techniques. Immunolocalisation studies revealed that the extracellular matrix layer outside the outer epidermal cell wall of B. napus embryogenic tissues contained napin. This is the first study to report the extracellular localisation of napin. In addition, we have also further characterised the expression pattern of Eg1 that encodes for napin in the B. napus embryogenic tissue.

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The Biosynthesis, Structure and Properties of Napin - the Storage Protein From Rape Seeds

Cited by 21 publications

References 50 publications

Large scale purification of rapeseed proteins (Brassica napus L.)

Large scale purification of rapeseed proteins (Brassica napus L.)

Recombinant pronapin precursor produced in Pichia pastoris displays structural and immunologic equivalent properties to its mature product isolated from rapeseed

Extracellular localization of napin in the embryogenic tissues of Brassica napus spp. oleifera

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