PXD013227

PXD013227 is an original dataset announced via ProteomeXchange.

Title	Deciphering mechanism of actions of novel compounds by bioactive thermal proteome profiling
Description	Exploring the therapeutic effects of bioactive compounds has been traditionally approaching by phenotypic screenings followed by functional validation by in vitro assays. The thermal proteome profiling (TPP) has been successfully applied to study drug targets and off-target. We applied a modified protocol based on TPP to elucidate the mechanism of actions (MOA)s of novel bioactives compounds from marine biodiscovery. We have modified the method to gain the specificity for its application to compounds with limited chemical or structural characterization. Method implementation includes increasing the centrifugation force to precipitate the microsomal fraction previous to the thermal shift assay. The range of temperatures has been reduced to optimize the analysis without compromising resolution. Finally, the mass spectrometry analysis was based on label-free quantitative proteomics. Comparison of the targets among methodologies confirmed that the precipitation of the microsomal membranes before TPP is an essential step to discriminate between true targets from proteins precipitates after subcellular fractionation by centrifugation force. As a probe of concept, a novel marine bioactive compound has been analyzed on the hepatic cell line HepG2. We found that 2 targets proteins, aldehyde dehydrogenase and isocitrate dehydoregenase, can be related with beneficial properties towards obesity and obesity-related comorbidities. Identification of targets is key to decipher the MOAs of bioactive compound, predict the mode of action as well as possible harmful effects.
HostingRepository	PRIDE
AnnounceDate	2019-06-28
AnnouncementXML	Submission_2019-06-28_00:35:28.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Ana Maria Carrasco del Amor
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	iodoacetamide derivatized residue
Instrument	LTQ Velos ETD

Revision	Datetime	Status	ChangeLog Entry
0	2019-03-25 07:21:31	ID requested
⏵ 1	2019-06-28 00:35:30	announced

Carrasco Del Amor A, Freitas S, Urbatzka R, Fresnedo O, Cristobal S, -Hydroxy-pheophytin Isolated from a Marine Cyanobacteria. Mar Drugs, 17(6):(2019) [pubmed]

submitter keyword: thermal proteome profiling

mechanism of actions

bioactive compound

label-free quantitative proteomics

marine biodiscovery

Susana Cristobal
contact affiliation	Department of Clinical and Experimental Medicine, Cell Biology, Faculty of Medicine, Linköping University, Linköping, Sweden. Department of Physiology, Ikerbasque, Faculty of Medicine and Nursing, University of the Basque Country UPV/EHU, Spain
contact email	susana.cristobal@liu.se
lab head
Ana Maria Carrasco del Amor
contact affiliation	Linköping University
contact email	ana.carrasco@liu.se
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/2019/06/PXD013227

PRIDE project URI

• PRIDE
  1. PXD013227
     1. Label: PRIDE project
     2. Name: Deciphering mechanism of actions of novel compounds by bioactive thermal proteome profiling