| Simulated gastrointestinal digestion of Pru ar 3 apricot allergen: Assessment of allergen resistance and characterization of the peptides by ultra-performance liquid chromatography/electrospray ionisation mass spectrometry. | |
| | |
MedLine Citation:
|
PMID: 23136021 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
|
RATIONALE: Non-specific lipid transfer proteins (ns-LTPs) are major food allergens of the Rosaceae family. The severity of allergic reactions often relates to resistance of the allergen to digestion. Thus, it is important to evaluate the digestibility of these proteins and characterise the peptides generated in the gastrointestinal tract. METHODS: Simulated gastrointestinal digestion of purified allergen Pru ar 3 was performed using pepsin for the gastric phase in aqueous HCl at pH = 2 and chymotrypsin and trypsin for the intestinal phase in aqueous NH(4) HCO(3) at pH = 7.8. The peptide mixture obtained was analysed by ultra-performance liquid chromatography/electrospray ionisation mass spectrometry (UPLC/ESI-MS). Peptide sequences were identified by comparing their molecular mass to that obtained by in silico digestion, and were confirmed by the ions obtained by in-source fragmentation. Semi-quantification was performed for the intact protein by comparison with internal standards. RESULTS: The resistance to gastrointestinal digestion of Pru ar 3 allergen was evaluated to be 9%. This value is consistent with that found for grape LTP, but much lower than the resistance found for peach LTP (35%). All the peptides generated were identified by ESI-MS on the basis of their molecular mass and from the ions generated from in-source fragmentation. Apart from low molecular mass peptides, five high molecular mass peptides (4500-7000 Da) containing disulphide bridges were identified. ESI-MS of the intact protein indicated a less compact folded structure when compared to that of the homologous peach LTP. CONCLUSIONS: An extensive characterisation of the peptides generated from the gastrointestinal digestion of Pru ar 3 allergen was performed here for the first time via UPLC/ESI-MS analysis. The digestibility of the allergen was evaluated and compared with that of other LTPs, demonstrating that only a small amount of undigested protein remains, and that specific proteolytic action involves immunodominant epitopes. These data might explain the lower allergenicity of apricot LTP compared to peach LTP, despite their high sequence homology. Copyright © 2012 John Wiley & Sons, Ltd. |
| | |
Authors:
|
Barbara Prandi; Laura Farioli; Tullia Tedeschi; Elide Anna Pastorello; Stefano Sforza |
Publication Detail:
|
Type: Journal Article |
Journal Detail:
|
Title: Rapid communications in mass spectrometry : RCM Volume: 26 ISSN: 1097-0231 ISO Abbreviation: Rapid Commun. Mass Spectrom. Publication Date: 2012 Dec |
Date Detail:
|
Created Date: 2012-11-08 Completed Date: - Revised Date: - |
Medline Journal Info:
|
Nlm Unique ID: 8802365 Medline TA: Rapid Commun Mass Spectrom Country: England |
Other Details:
|
Languages: eng Pagination: 2905-12 Citation Subset: IM |
Copyright Information:
|
Copyright © 2012 John Wiley & Sons, Ltd. |
Affiliation:
|
Department of Food Science, University of Parma, viale delle Scienze 17a, I-43124, Parma, Italy. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
|
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Differentiation of isomeric pentose disaccharides by electrospray ionization tandem mass spectrometr...
Next Document: Identification of xanthones from Swertia punicea using high-performance liquid chromatography couple...