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dc.creatorNešić, Andrijana
dc.creatorČavić, Milena
dc.creatorPopović, Milica
dc.creatorGavrović-Jankulović, Marija
dc.date.accessioned2021-09-02T10:49:51Z
dc.date.available2021-09-02T10:49:51Z
dc.date.issued2021
dc.identifier.issn0301-4851
dc.identifier.urihttps://cherry.chem.bg.ac.rs/handle/123456789/4549
dc.description.abstractActinidin (Act d 1), a highly abundant cysteine protease from kiwifruit, is one of the major contributors to the development of kiwifruit allergy. Many studies have focused on the optimization of Act d 1 purification and its role in the development of food allergies. Testing on cell culture monolayers is a common step in the elucidation of food allergen sensitization. In the case of cysteine proteases, an additional activation step with l-cysteine is required before the testing. Hence, we aimed to evaluate whether l-cysteine already present in commonly used cell culture media would suffice for Act d 1 activation. Successfully activated Act d 1 (98.1% of proteolytic activity, as compared to l-cysteine activated Act d 1) was further tested in two commonly used 2D model systems (Caco-2 and HEK293 cells) to evaluate its role on the mRNA expression of cytokines involved in the innate immunity (IL-1β, IL-6, TNFα, TSLP). Furthermore, the contribution of Act d 1 in the promotion of inflammation through regulation of inducible nitric oxide synthase (iNOS) mRNA expression was also examined. These results demonstrate that activation of cysteine proteases can be achieved without previous enzyme incubation in l-cysteine -containing solution. Act d 1 incubated in cell culture medium was able to modulate gene expression of pro-inflammatory cytokines when tested on two model systems of the epithelial barrier. © 2021, The Author(s), under exclusive licence to Springer Nature B.V.
dc.languageEnglish
dc.publisherSpringer Nature
dc.rightsrestrictedAccess
dc.sourceMolecular Biology Reports
dc.subjectActinidin
dc.subjectCysteine protease
dc.subjectKiwifruit allergen
dc.subjectPro-inflammatory cytokines
dc.titleA new approach for activation of the kiwifruit cysteine protease for usage in in-vitro testing
dc.typearticleen
dc.rights.licenseARR
dcterms.abstractПоповић, Милица; Гавровић-Јанкуловић, Марија; Нешић, Aндријана; Чавић, Милена;
dc.citation.volume48
dc.citation.issue5
dc.citation.spage4065
dc.citation.epage4072
dc.identifier.wos000659391800003
dc.identifier.doi10.1007/s11033-021-06416-8
dc.citation.rankM23~
dc.type.versionpublishedVersion
dc.identifier.scopus2-s2.0-85107467674


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