PXD010634

PXD010634 is an original dataset announced via ProteomeXchange.

Title	Profiling N-myristoyltransferase activity and inhibition in breast cancer cells (MD-MB-231) using Sortase-A labelling
Description	We report a novel application for Sortase A (SrtA), labeling N-terminal Gly proteins across the proteome at endogenous levels. We combined SrtA labeling of whole-cell lysates with a potent and selective NMT inhibitor, delivering a method capable of quantifying changes in cellular NMT activity in multiple substrates at the whole proteome level. This method shows good overlap in substrates with a previously reported YnMyr metabolic tagging strategy (PXD002687), and a similar capability to measure cellular NMT activity. The SrtA-based approach described here is complementary to YnMyr metabolic tagging, since the increase in signal for newly synthesized proteins exposing an N-terminal glycine mirrors the loss of signal seen with YnMyr. Importantly, this new method overcomes the limitations imposed by metabolic tagging. Since SrtA labeling is performed post-lysis this labeling strategy provides a readout of the cell status in the absence of any external perturbation and is applicable to virtually any type of biological sample, giving clear advantages over other in-cell labeling strategies. We anticipate that this method will enable the exploration of new avenues for NMT as a therapeutic target by providing a method for de novo identification of substrates most responsive to NMT inhibition, and a multifuctional target engagement biomarker assay suited to both animal models and patient biopsies. Besides shotgun proteomics, we coupled this robust NMT activity assay to a variety of other detection techniques, including gel-based analyses, and ELISA. This submission concerns myristoylated proteomes of human origin.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_04:47:31.656.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Remigiusz Serwa
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	myristoylated residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2018-07-31 07:39:28	ID requested
1	2018-10-25 09:13:37	announced
⏵ 2	2024-10-22 04:47:32	announced	2024-10-22: Updated project metadata.

Goya Grocin A, Serwa RA, Morales Sanfrutos J, Ritzefeld M, Tate EW, -Myristoyltransferase Activity and Inhibition Using Sortase A. Mol Cell Proteomics, 18(1):115-126(2019) [pubmed]

10.1074/mcp.ra118.001043;

curator keyword: Biological, Biomedical

submitter keyword: Sortase A

SrtA

N-myristoylation

NMT

Edward W Tate
contact affiliation	Institute of Chemical Biology, Department of Chemistry, Imperial College London
contact email	e.tate@imperial.ac.uk
lab head
Remigiusz Serwa
contact affiliation	Imperial College London
contact email	r.serwa@imperial.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD010634
     1. Label: PRIDE project
     2. Name: Profiling N-myristoyltransferase activity and inhibition in breast cancer cells (MD-MB-231) using Sortase-A labelling