PXD012156

PXD012156 is an original dataset announced via ProteomeXchange.

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

Revision	Datetime	Status	ChangeLog Entry
0	2018-12-21 05:50:23	ID requested
⏵ 1	2019-03-18 08:21:57	announced

H, ö, 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]

submitter keyword: optogenetics, affinity purification, phytochrome, ZAP70, protein-protein interaction, mass spectrometry

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

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PRIDE project URI

• PRIDE
  1. PXD012156
     1. Label: PRIDE project
     2. Name: Light-controlled affinity purification of protein complexes