Proteolysis of the peanut allergen Ara h 1 by an endogenous aspartic protease
Copyright © 2015 Elsevier Masson SAS. All rights reserved.
Veröffentlicht in: | Plant physiology and biochemistry : PPB. - 1991. - 96(2015) vom: 27. Nov., Seite 301-10 |
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Format: | Online-Aufsatz |
Sprache: | English |
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2015
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Zugriff auf das übergeordnete Werk: | Plant physiology and biochemistry : PPB |
Schlagworte: | Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. 7S globulin storage protein Allergen Ara h 1 Aspartic protease Germination Peanut Proteolysis mehr... |
Zusammenfassung: | Copyright © 2015 Elsevier Masson SAS. All rights reserved. The 7S and 11S globulins of peanuts are subjected to proteolysis two days after seed imbibition, with Ara h 1 and the arachin acidic chains being among the first storage proteins to be mobilized. Proteolytic activity was greatest at pH 2.6-3 and is inhibited by pepstatin A, characteristic of an aspartic protease. This activity persists in seedling cotyledons up to at least 8 days after imbibition. In vitro proteolysis of Ara h 1 at pH 2.6 by extracts of cotyledons from seedlings harvested 24 h after seed imbibition generates newly appearing bands on SDS-PAGE. Partial sequences of Ara h 1 that were obtained through LC-MS/MS analysis of in-gel trypsin digests of those bands, combined with information on fragment size, suggest that proteolysis begins in the region that links the two cupin domains to produce two 33/34 kD fragments, each one encompassing an intact cupin domain. The later appearance of two 18 and 10/11 kD fragments can be explained by proteolysis within an exposed site in the cupin domains of each of the 33/34 kD fragments. The same or similar proteolytic activity was observed in developing seeds, but Ara h 1 remains intact through seed maturation. This is partly explained by the observation that acidification of the protein storage vacuoles, demonstrated by vacuolar accumulation of acridine orange that was dissipated by a membrane-permeable base, occurs only after germination. These findings suggest a method for use of the seed aspartic protease in reducing peanut allergy due to Ara h 1 |
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Beschreibung: | Date Completed 02.09.2016 Date Revised 30.09.2020 published: Print-Electronic Citation Status MEDLINE |
ISSN: | 1873-2690 |
DOI: | 10.1016/j.plaphy.2015.08.008 |