Please use this identifier to cite or link to this item: http://dx.doi.org/10.14279/depositonce-11521
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Main Title: Quantitative photo-crosslinking mass spectrometry revealing protein structure response to environmental changes
Author(s): Müller, Fränze
Graziadei, Andrea
Rappsilber, Juri
Type: Article
URI: https://depositonce.tu-berlin.de/handle/11303/12721
http://dx.doi.org/10.14279/depositonce-11521
License: https://creativecommons.org/licenses/by/4.0/
Abstract: Protein 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.
Subject(s): peptides
proteins
monomers
pH
cluster chemistry
conformational transitions
Issue Date: 17-Jun-2019
Date Available: 4-Mar-2021
Is Part Of: 10.14279/depositonce-9814
Language Code: en
DDC Class: 540 Chemie und zugeordnete Wissenschaften
Journal Title: Analytical chemistry
Publisher: American Chemical Society (ACS)
Volume: 91
Issue: 14
Publisher DOI: 10.1021/acs.analchem.9b01339
Page Start: 9041
Page End: 9048
EISSN: 1520-6882
ISSN: 0003-2700
TU Affiliation(s): Fak. 3 Prozesswissenschaften » Inst. Biotechnologie
Appears in Collections:Technische Universität Berlin » Publications

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