Please use this identifier to cite or link to this item: http://dx.doi.org/10.14279/depositonce-11521
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dc.contributor.authorMüller, Fränze-
dc.contributor.authorGraziadei, Andrea-
dc.contributor.authorRappsilber, Juri-
dc.date.accessioned2021-03-04T09:59:39Z-
dc.date.available2021-03-04T09:59:39Z-
dc.date.issued2019-06-17-
dc.identifier.issn0003-2700-
dc.identifier.urihttps://depositonce.tu-berlin.de/handle/11303/12721-
dc.identifier.urihttp://dx.doi.org/10.14279/depositonce-11521-
dc.description.abstractProtein structures respond to changes in their chemical and physical environment. However, studying such conformational changes is notoriously difficult, as many structural biology techniques are also affected by these parameters. Here, the use of photo-crosslinking, coupled with quantitative crosslinking mass spectrometry (QCLMS), offers an opportunity, since the reactivity of photo-crosslinkers is unaffected by changes in environmental parameters. In this study, we introduce a workflow combining photo-crosslinking using sulfosuccinimidyl 4,4′-azipentanoate (sulfo-SDA) with our recently developed data-independent acquisition (DIA)-QCLMS. This novel photo-DIA-QCLMS approach is then used to quantify pH-dependent conformational changes in human serum albumin (HSA) and cytochrome C by monitoring crosslink abundances as a function of pH. Both proteins show pH-dependent conformational changes resulting in acidic and alkaline transitions. 93% and 95% of unique residue pairs (URP) were quantifiable across triplicates for HSA and cytochrome C, respectively. Abundance changes of URPs and hence conformational changes of both proteins were visualized using hierarchical clustering. For HSA we distinguished the N–F and the N–B form from the native conformation. In addition, we observed for cytochrome C acidic and basic conformations. In conclusion, our photo-DIA-QCLMS approach distinguished pH-dependent conformers of both proteins.en
dc.language.isoenen
dc.relation.ispartof10.14279/depositonce-9814-
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/en
dc.subject.ddc540 Chemie und zugeordnete Wissenschaftende
dc.subject.otherpeptidesen
dc.subject.otherproteinsen
dc.subject.othermonomersen
dc.subject.otherpHen
dc.subject.othercluster chemistryen
dc.subject.otherconformational transitionsen
dc.titleQuantitative photo-crosslinking mass spectrometry revealing protein structure response to environmental changesen
dc.typeArticleen
tub.accessrights.dnbfreeen
tub.publisher.universityorinstitutionTechnische Universität Berlinen
dc.identifier.eissn1520-6882-
dc.type.versionpublishedVersionen
dcterms.bibliographicCitation.doi10.1021/acs.analchem.9b01339en
dcterms.bibliographicCitation.journaltitleAnalytical chemistryen
dcterms.bibliographicCitation.originalpublisherplaceColumbus, Ohioen
dcterms.bibliographicCitation.volume91en
dcterms.bibliographicCitation.pageend9048en
dcterms.bibliographicCitation.pagestart9041en
dcterms.bibliographicCitation.originalpublishernameAmerican Chemical Society (ACS)en
dcterms.bibliographicCitation.issue14en
tub.affiliationFak. 3 Prozesswissenschaften » Inst. Biotechnologiede
Appears in Collections:Technische Universität Berlin » Publications

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