Determination of Allergen Levels, Isoforms, and Their Hydroxyproline Modifications Among Peanut Genotypes by Mass Spectrometry

The recently published reference genome of peanuts enables a detailed molecular description of the allergenic proteins of the seed. We used LC-MS/MS to investigate peanuts of different genotypes to assess variability and to better describe naturally occurring allergens and isoforms. Using relative q...

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Main Authors: Justin T. Marsh, Lee K. Palmer, Stef J. Koppelman, Philip E. Johnson
Format: Article
Language:English
Published: Frontiers Media S.A. 2022-05-01
Series:Frontiers in Allergy
Subjects:
Online Access:https://www.frontiersin.org/articles/10.3389/falgy.2022.872714/full
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author Justin T. Marsh
Lee K. Palmer
Stef J. Koppelman
Philip E. Johnson
author_facet Justin T. Marsh
Lee K. Palmer
Stef J. Koppelman
Philip E. Johnson
author_sort Justin T. Marsh
collection DOAJ
description The recently published reference genome of peanuts enables a detailed molecular description of the allergenic proteins of the seed. We used LC-MS/MS to investigate peanuts of different genotypes to assess variability and to better describe naturally occurring allergens and isoforms. Using relative quantification by mass spectrometry, minor variation of some allergenic proteins was observed, but total levels of Ara h 1, 2, 3, and 6 were relatively consistent among 20 genotypes. Previously published RP-HPLC methodology was used for comparison. The abundance of three Ara h 3 isoforms were variable among the genotypes and contributed to a large proportion of total Ara h 3 where present. Previously unpublished hydroxyproline sites were identified in Ara h 1 and 3. Hydroxylation did not vary significantly where sites were present. Peanut allergen composition was largely stable, with only some isoforms displaying differences between genotypes. The resulting differences in allergenicity are of unknown clinical significance but are likely to be minor. The data presented herein allow for the design of targeted MS methodology to allow the quantitation and therefore control of peanut allergens of clinical relevance and observed variability.
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spelling doaj.art-62bc5df42f6946d9b0686adbe3b92eab2022-12-22T03:23:10ZengFrontiers Media S.A.Frontiers in Allergy2673-61012022-05-01310.3389/falgy.2022.872714872714Determination of Allergen Levels, Isoforms, and Their Hydroxyproline Modifications Among Peanut Genotypes by Mass SpectrometryJustin T. MarshLee K. PalmerStef J. KoppelmanPhilip E. JohnsonThe recently published reference genome of peanuts enables a detailed molecular description of the allergenic proteins of the seed. We used LC-MS/MS to investigate peanuts of different genotypes to assess variability and to better describe naturally occurring allergens and isoforms. Using relative quantification by mass spectrometry, minor variation of some allergenic proteins was observed, but total levels of Ara h 1, 2, 3, and 6 were relatively consistent among 20 genotypes. Previously published RP-HPLC methodology was used for comparison. The abundance of three Ara h 3 isoforms were variable among the genotypes and contributed to a large proportion of total Ara h 3 where present. Previously unpublished hydroxyproline sites were identified in Ara h 1 and 3. Hydroxylation did not vary significantly where sites were present. Peanut allergen composition was largely stable, with only some isoforms displaying differences between genotypes. The resulting differences in allergenicity are of unknown clinical significance but are likely to be minor. The data presented herein allow for the design of targeted MS methodology to allow the quantitation and therefore control of peanut allergens of clinical relevance and observed variability.https://www.frontiersin.org/articles/10.3389/falgy.2022.872714/fullArachis hypogeaAra h 1Ara h 2Ara h 3Ara h 6food allergy
spellingShingle Justin T. Marsh
Lee K. Palmer
Stef J. Koppelman
Philip E. Johnson
Determination of Allergen Levels, Isoforms, and Their Hydroxyproline Modifications Among Peanut Genotypes by Mass Spectrometry
Frontiers in Allergy
Arachis hypogea
Ara h 1
Ara h 2
Ara h 3
Ara h 6
food allergy
title Determination of Allergen Levels, Isoforms, and Their Hydroxyproline Modifications Among Peanut Genotypes by Mass Spectrometry
title_full Determination of Allergen Levels, Isoforms, and Their Hydroxyproline Modifications Among Peanut Genotypes by Mass Spectrometry
title_fullStr Determination of Allergen Levels, Isoforms, and Their Hydroxyproline Modifications Among Peanut Genotypes by Mass Spectrometry
title_full_unstemmed Determination of Allergen Levels, Isoforms, and Their Hydroxyproline Modifications Among Peanut Genotypes by Mass Spectrometry
title_short Determination of Allergen Levels, Isoforms, and Their Hydroxyproline Modifications Among Peanut Genotypes by Mass Spectrometry
title_sort determination of allergen levels isoforms and their hydroxyproline modifications among peanut genotypes by mass spectrometry
topic Arachis hypogea
Ara h 1
Ara h 2
Ara h 3
Ara h 6
food allergy
url https://www.frontiersin.org/articles/10.3389/falgy.2022.872714/full
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