PXD022797

PXD022797 is an original dataset announced via ProteomeXchange.

Title	Design of a proteolytically stable sodium-calcium exchanger 1 (NCX1)-phospholemman (PLM) disruptor peptide for in vivo studies
Description	The cardiac sodium–calcium exchanger (NCX1) is important for normal Na+ and Ca2+ homeostasis and cardiomyocyte relaxation and contraction. NCX1 activity is reduced by its endogenous inhibitor phosphorylated phospholemman (pSer68-PLM). Disrupting the pSer68-PLM-NCX1 interaction is therefore a potential therapeutic strategy to reverse NCX1 inhibition in cardiac disease. We have previously designed a high affinity NCX1-PLM disruptor peptide (OPT) that reverses the inhibitory pSer68-PLM-NCX1 interaction in HEK293 cells. In the present study, we performed N and C-terminal truncation analyses of OPT and identified PYKEIEQLIELANYQV as the minimal pSer68-PLM binding sequence. To increase peptide stability in human serum, we replaced the proline in its N-terminus with an N-methyl-proline (NOPT) after identification of N-terminus as substitution tolerant by two dimensional peptide array analysis. Mass spectrometry analysis revealed that the half-life of the novel NOPT peptide was increased 10-fold compared to that of OPT. Biotinylated NOPT pulled down endogenous PLM from rat left ventricle lysate and exhibited pSer68-PLM binding in an ELISA-based assay. Importantly, NOPT reduced the PLM-NCX1 interaction in an ELISA-based assay and cell permeable NOPT-TAT increased the NCX1 activity in adult cardiomyocytes isolated from both wild type (WT) and HF animals. Finally, we found that NOPT interacted with pSer68-PLM with a KD value of 2.7±3.8 μM, whereas the NCX1 cytoplasmic part interacted with PLM and pSer68-PLM with KD values of 14±2 nM and 24±14 nM. In conclusion, we have here developed a proteolytically stable NCX1-PLM disruptor peptide that upregulates NCX1 activity in WT and HF cardiomyocytes.
HostingRepository	PRIDE
AnnounceDate	2021-09-09
AnnouncementXML	Submission_2021-09-09_14:48:01.683.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Tuula Nyman
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2020-11-27 08:49:17	ID requested
⏵ 1	2021-09-09 14:48:02	announced

Wanichawan P, Skogestad J, Lunde M, St, ø, le TP, Stensland M, Nyman TA, Sjaastad I, Sejersted OM, Aronsen JM, Carlson CR, Studies. Front Pharmacol, 12():638646(2021) [pubmed]

submitter keyword: phospholemman

sodium-calcium exchanger

peptidomimetics

peptide array

cardiac

Tuula Nyman
contact affiliation	Head of Proteomics
contact email	tuula.nyman@medisin.uio.no
lab head
Tuula Nyman
contact affiliation	University of Oslo
contact email	tuula.nyman@medisin.uio.no
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD022797
     1. Label: PRIDE project
     2. Name: Design of a proteolytically stable sodium-calcium exchanger 1 (NCX1)-phospholemman (PLM) disruptor peptide for in vivo studies