PXD018047

PXD018047 is an original dataset announced via ProteomeXchange.

Title	Tissue-specific protein complex analysis with natively processed bait proteins
Description	Protein-protein interaction analysis is an important tool to elucidate the function of proteins and protein complexes as well as their dynamic behavior. Despite the large toolset available to date the analysis of tissue specific complexes is still relying on the availability of specific antibodies suited for immunoprecipitation. Due to the lack of specific, high affinity antibodies, we aimed at establishing a novel approach that is generally applicable to identify protein interactions and complexes in tissue. Therefore, we established an approach for which a tagged bait protein is expressed in a mammalian cell line which is, after a specific cleaning procedure, used to pulldown prey proteins and complexes from tissue samples. We demonstrate here that this protocol allows the identification of tissue specific interaction by applying it to the ciliary protein lebercilin and its specific interactors were identified in retina or the retinal pigment epithelium. Further, the heart specific interactome of two isoforms of the cyclic guanosine-3’,5’-monophosphate (cGMP)-dependent protein kinase type 1α and 1β (cGKI/ aka PRKG1/), which differ only in their unique amino-terminal region comprising about 100 AS, shows the suitability of the approach to identify isoform-specific interactions. With this affinity purification-based protocol, we provide a fast and reliable method for tissue-specific protein complex analysis independent of antibody availability.
HostingRepository	PRIDE
AnnounceDate	2023-12-28
AnnouncementXML	Submission_2023-12-28_13:11:45.055.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Tina Beyer
SpeciesList	scientific name: Sus scrofa domesticus (domestic pig); NCBI TaxID: 9825;
ModificationList	carbamoylated residue; acetylated residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2020-03-13 04:59:02	ID requested
⏵ 1	2023-12-28 13:11:45	announced

10.1016/j.jprot.2020.103947;

Beyer T, Klose F, Kuret A, Hoffmann F, Lukowski R, Ueffing M, Boldt K, Tissue- and isoform-specific protein complex analysis with natively processed bait proteins. J Proteomics, 231():103947(2021) [pubmed]

submitter keyword: tissue-specific protein complex analysis,Isoform-specific

Marius Ueffing
contact affiliation	Institute for Ophthalmic Research, University Clinics Tuebingen
contact email	marius.ueffing@uni-tuebingen.de
lab head
Tina Beyer
contact affiliation	University Clinics Tuebingen
contact email	tina.beyer@uni-tuebingen.de
dataset submitter

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/2023/12/PXD018047

PRIDE project URI

• PRIDE
  1. PXD018047
     1. Label: PRIDE project
     2. Name: Tissue-specific protein complex analysis with natively processed bait proteins