Gastric digestome of whole peanut grains from the aspect of immunoproteomics: Characterization of digested allergens in the real food matrix
Аутори
Prodić, IvanaStanić-Vučinić, Dragana
Apostolović, Danijela
Radosavljević, Jelena
Mihailović, Jelena
Smiljanić, Katarina
Ćirković-Veličković, Tanja
Конференцијски прилог (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
Objective: Major peanut allergens, Ara h 2 and Ara h 6, are known to be resistant to pepsindigestion, and they sensitize individual via the gastrointestinal tract. Mikenus et al. published astandardized static digestion method for food, based on physiological conditions emphasizing theimpact of food matrices. Immunoreactive proteins (large fragments) and peptides (short digestionresistant peptides SDRPs; <10 kDa), to which the immune system of the gastrointestinal tract isexposed during digestion of peanut proteins, has not been investigated under pure physiologicalconditions suggested by this protocol.Matherial and methods: Whole grain of grounded raw peanut was incubated with human α-amylase, and pepsin, mimicking the effects of oral and gastric digestion, in total duration of 2h.Bottom up proteomic approach, immunoblotting with allergen-specific antibodies from peanut-sensitized patients, enzyme-linked immunosorbent inhibition assay and ImmunoCAP tests, wereused to identify and char...acterize peanut digesta.Results: After 2h of oral/gastric phase we got, intact proteins, large, digestion resistant peptides(DRP) and SDRPs, as well. Ara h 2 and Ara h 6 remained mostly intact, and short DRPs from Ara h2 and Ara h 6 were more potent in inhibiting IgE binding than Ara h 1 and Ara 3. Ara h 1 and Ara h3 showed preserved allergenic capacity, as well. Almost all of identified short DRPs from Ara h 1,Ara h 2 and Ara h 3, with preserved allergenic potential, were constituents of continuous epitopesequences found via Immune Epitope Database (www.iedb.org).Conclusion: Processes of protein extraction from the matrix and their enzymatic digestion occursimultaneously. Oral and gastric phase digestion products of raw peanut are intact proteins, largeand short digestion resistant peptides. Under these conditions Ara h 2 and Ara h 6 are expectedly
Кључне речи:
digestomics / peanut allergens / gastric digestion / INFOGEST 1.0Извор:
IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija, 2018Издавач:
- Srpsko Udruženje za Proteomiku; IBISS
Финансирање / пројекти:
- Молекуларне особине и модификације неких респираторних и нутритивних алергена (RS-MESTD-Basic Research (BR or ON)-172024)
Напомена:
- Book of Abstracts
Институција/група
TorlakTY - CONF AU - Prodić, Ivana AU - Stanić-Vučinić, Dragana AU - Apostolović, Danijela AU - Radosavljević, Jelena AU - Mihailović, Jelena AU - Smiljanić, Katarina AU - Ćirković-Veličković, Tanja PY - 2018 UR - http://intor.torlakinstitut.com/handle/123456789/789 AB - Objective: Major peanut allergens, Ara h 2 and Ara h 6, are known to be resistant to pepsindigestion, and they sensitize individual via the gastrointestinal tract. Mikenus et al. published astandardized static digestion method for food, based on physiological conditions emphasizing theimpact of food matrices. Immunoreactive proteins (large fragments) and peptides (short digestionresistant peptides SDRPs; <10 kDa), to which the immune system of the gastrointestinal tract isexposed during digestion of peanut proteins, has not been investigated under pure physiologicalconditions suggested by this protocol.Matherial and methods: Whole grain of grounded raw peanut was incubated with human α-amylase, and pepsin, mimicking the effects of oral and gastric digestion, in total duration of 2h.Bottom up proteomic approach, immunoblotting with allergen-specific antibodies from peanut-sensitized patients, enzyme-linked immunosorbent inhibition assay and ImmunoCAP tests, wereused to identify and characterize peanut digesta.Results: After 2h of oral/gastric phase we got, intact proteins, large, digestion resistant peptides(DRP) and SDRPs, as well. Ara h 2 and Ara h 6 remained mostly intact, and short DRPs from Ara h2 and Ara h 6 were more potent in inhibiting IgE binding than Ara h 1 and Ara 3. Ara h 1 and Ara h3 showed preserved allergenic capacity, as well. Almost all of identified short DRPs from Ara h 1,Ara h 2 and Ara h 3, with preserved allergenic potential, were constituents of continuous epitopesequences found via Immune Epitope Database (www.iedb.org).Conclusion: Processes of protein extraction from the matrix and their enzymatic digestion occursimultaneously. Oral and gastric phase digestion products of raw peanut are intact proteins, largeand short digestion resistant peptides. Under these conditions Ara h 2 and Ara h 6 are expectedly PB - Srpsko Udruženje za Proteomiku; IBISS C3 - IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija T1 - Gastric digestome of whole peanut grains from the aspect of immunoproteomics: Characterization of digested allergens in the real food matrix UR - https://hdl.handle.net/21.15107/rcub_intor_789 ER -
@conference{ author = "Prodić, Ivana and Stanić-Vučinić, Dragana and Apostolović, Danijela and Radosavljević, Jelena and Mihailović, Jelena and Smiljanić, Katarina and Ćirković-Veličković, Tanja", year = "2018", abstract = "Objective: Major peanut allergens, Ara h 2 and Ara h 6, are known to be resistant to pepsindigestion, and they sensitize individual via the gastrointestinal tract. Mikenus et al. published astandardized static digestion method for food, based on physiological conditions emphasizing theimpact of food matrices. Immunoreactive proteins (large fragments) and peptides (short digestionresistant peptides SDRPs; <10 kDa), to which the immune system of the gastrointestinal tract isexposed during digestion of peanut proteins, has not been investigated under pure physiologicalconditions suggested by this protocol.Matherial and methods: Whole grain of grounded raw peanut was incubated with human α-amylase, and pepsin, mimicking the effects of oral and gastric digestion, in total duration of 2h.Bottom up proteomic approach, immunoblotting with allergen-specific antibodies from peanut-sensitized patients, enzyme-linked immunosorbent inhibition assay and ImmunoCAP tests, wereused to identify and characterize peanut digesta.Results: After 2h of oral/gastric phase we got, intact proteins, large, digestion resistant peptides(DRP) and SDRPs, as well. Ara h 2 and Ara h 6 remained mostly intact, and short DRPs from Ara h2 and Ara h 6 were more potent in inhibiting IgE binding than Ara h 1 and Ara 3. Ara h 1 and Ara h3 showed preserved allergenic capacity, as well. Almost all of identified short DRPs from Ara h 1,Ara h 2 and Ara h 3, with preserved allergenic potential, were constituents of continuous epitopesequences found via Immune Epitope Database (www.iedb.org).Conclusion: Processes of protein extraction from the matrix and their enzymatic digestion occursimultaneously. Oral and gastric phase digestion products of raw peanut are intact proteins, largeand short digestion resistant peptides. Under these conditions Ara h 2 and Ara h 6 are expectedly", publisher = "Srpsko Udruženje za Proteomiku; IBISS", journal = "IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija", title = "Gastric digestome of whole peanut grains from the aspect of immunoproteomics: Characterization of digested allergens in the real food matrix", url = "https://hdl.handle.net/21.15107/rcub_intor_789" }
Prodić, I., Stanić-Vučinić, D., Apostolović, D., Radosavljević, J., Mihailović, J., Smiljanić, K.,& Ćirković-Veličković, T.. (2018). Gastric digestome of whole peanut grains from the aspect of immunoproteomics: Characterization of digested allergens in the real food matrix. in IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija Srpsko Udruženje za Proteomiku; IBISS.. https://hdl.handle.net/21.15107/rcub_intor_789
Prodić I, Stanić-Vučinić D, Apostolović D, Radosavljević J, Mihailović J, Smiljanić K, Ćirković-Veličković T. Gastric digestome of whole peanut grains from the aspect of immunoproteomics: Characterization of digested allergens in the real food matrix. in IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija. 2018;. https://hdl.handle.net/21.15107/rcub_intor_789 .
Prodić, Ivana, Stanić-Vučinić, Dragana, Apostolović, Danijela, Radosavljević, Jelena, Mihailović, Jelena, Smiljanić, Katarina, Ćirković-Veličković, Tanja, "Gastric digestome of whole peanut grains from the aspect of immunoproteomics: Characterization of digested allergens in the real food matrix" in IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija (2018), https://hdl.handle.net/21.15107/rcub_intor_789 .