PXD031003

PXD031003 is an original dataset announced via ProteomeXchange.

Title	Mapping of phosphorylation target sites of obscurin-kin 1 within the cytoplasmic domain of N-cadherin by LC-MS/MS
Description	To test our hypothesis that obscurin-kin1 modulates, at least in part, the mechanochemical coupling of cardiomyocytes via phosphorylation of N-cadherin, which has been shown to play a pivotal role in these processes, we set forth to identify the potential phosphorylation target sites of obscurin-kin1 within the cytoplasmic domain of N-cadherin. To this end, we used the baculovirus system to produce catalytically active kin1 coupled to 6xHis-tag (His-Kin1-CA) and the bacterial system to produce an N-cadherin construct containing a portion of the cytoplasmic domain (aa 786-880, accession no. P15116) tagged to 6xHis (His-Ncad786-880), which is enriched in Ser residues predicted to be potent phosphorylation target sites by EXPASY/GPS2.1. Following affinity purification of the recombinant proteins, His-Ncad786-880 was subjected to an in vitro kinase assay in the presence of His-Kin1-CA or control baculovirus preparation infected with empty vector that had undergone the same purification process as His-Kin1-CA. The reaction mixtures were subsequently subjected to liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis. An N-cadherin peptide encompassing amino acids 786-807 containing a single phosphorylation event at Ser-788 was repeatedly identified in the His-Ncad786-880 reaction mixture when treated with His-Kin1-CA, but not in the control preparation
HostingRepository	PRIDE
AnnounceDate	2024-05-23
AnnouncementXML	Submission_2024-05-22_23:39:18.622.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Li Wang
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	phosphorylated residue
Instrument	LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2022-01-16 22:56:14	ID requested
⏵ 1	2024-05-22 23:39:19	announced

10.1172/jci.insight.162178;

Wang L, Tsakiroglou P, Gonzales R, Cho S, Li A, Dos Remedios C, Wright N, Kontrogianni-Konstantopoulos A, Essential role of obscurin kinase-1 in cardiomyocyte coupling via N-cadherin phosphorylation. JCI Insight, 9(3):(2024) [pubmed]

submitter keyword: cytoplasmic domain of N-cadherin,obscurin kinase 1, Ser phosphorylation, LC-MS/MS

Aikaterini Kontrogianni-Konstantopoulos
contact affiliation	Aikaterini Kontrogianni-Konstantopoulos, Ph.D. Professor of Biochemistry and Molecular Biology Director, Interdisciplinary Training Program in Muscle Biology Marlene and Stewart Greenebaum Comprehensive Cancer Center University of Maryland School of Medicine 108 N. Greene Street, 215A Baltimore, MD 21201
contact email	AKontrogianni@som.umaryland.edu
lab head
Li Wang
contact affiliation	​Scientist, HJF/Armed Forces Radiobiology Research Institute Assistant Professor (Adjunct)/Department of Pathology, USUHS
contact email	li.wang.ctr1@usuhs.edu
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/2024/05/PXD031003

PRIDE project URI

• PRIDE
  1. PXD031003
     1. Label: PRIDE project
     2. Name: Mapping of phosphorylation target sites of obscurin-kin 1 within the cytoplasmic domain of N-cadherin by LC-MS/MS