PXD012156 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Light-controlled affinity purification of protein complexes |
Description | Methods to affinity purify proteins are widely used in protein research. One important application is to identify interacting proteins of an affinity-purified protein of interest (POI) by mass spectrometry. Here, we developed an optogenetics-derived and light-controlled affinity purification method based on the light-regulated reversible protein interaction between phytochrome B (PhyB) and its phytochrome interacting factor 6 (PIF6). We engineered a truncated variant of PIF6 comprising only 22 amino acids that can be genetically fused to the POI as an affinity tag. Thereby the POI can be purified with PhyB-functionalized resin material using 660 nm light for binding and washing and 740 nm light for elution. As proof-of-concept, we expressed PIF-tagged variants of the tyrosine kinase ZAP70 in ZAP70-deficient Jurkat T cells, purified ZAP70 and associating proteins using our light-controlled system and identified the interaction partners by mass spectrometry |
HostingRepository | PRIDE |
AnnounceDate | 2019-03-18 |
AnnouncementXML | Submission_2019-03-18_08:21:56.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Jennifer Schwarz |
SpeciesList | scientific name: Homo sapiens (Human); NCBI TaxID: 9606; |
ModificationList | acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue |
Instrument | Q Exactive |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2018-12-21 05:50:23 | ID requested | |
⏵ 1 | 2019-03-18 08:21:57 | announced | |
Publication List
H, รถ, rner M, Eble J, Yousefi OS, Schwarz J, Warscheid B, Weber W, Schamel WWA, Light-Controlled Affinity Purification of Protein Complexes Exemplified by the Resting ZAP70 Interactome. Front Immunol, 10():226(2019) [pubmed] |
Keyword List
submitter keyword: optogenetics, affinity purification, phytochrome, ZAP70, protein-protein interaction, mass spectrometry |
Contact List
Bettina Warscheid |
contact affiliation | Faculty of Biology, University of Freiburg, Germany Signalling Research Centres BIOSS and CIBSS, University of Freiburg, Germany |
contact email | bettina.warscheid@biologie.uni-freiburg.de |
lab head | |
Jennifer Schwarz |
contact affiliation | University of Freiburg |
contact email | jennifer.schwarz@sgbm.uni-freiburg.de |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/03/PXD012156 |
PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD012156
- Label: PRIDE project
- Name: Light-controlled affinity purification of protein complexes