PXD017346

PXD017346 is an original dataset announced via ProteomeXchange.

Title	Selection of Heating Temperatures Improves the Sensitivity of Proteome Integral Solubility Alteration (PISA) Assay
Description	The thermal shift assay is a robust method of discovering protein-ligand interactions by measuring the alterations in protein thermal stability under various conditions. Several thermal shift assays have been developed and their throughput has been advanced greatly by the rapid progress in tandem mass tag-based quantitative proteomics. A recent paper by Gaetani et al. (J Proteome Res 2019, 18 (11), 4027-4037) introduced proteome integral solubility alteration (PISA) assay, further increasing throughput and simplifying the data analysis. Yet, it remains unclear how fold changes (integral treated samples versus integral control samples) perform in this assay. We show that fold changes have compromised linearity with ΔTm (shift in melting points) by simulation, and the magnitudes of the fold changes are inherently small in PISA assay, which is a challenge for quantitation. Both simulation and experimental results show that the selection of heating temperatures can tackle the small fold change problem and improve the sensitivity and specificity of the PISA assay.
HostingRepository	PRIDE
AnnounceDate	2020-04-06
AnnouncementXML	Submission_2020-04-30_03:35:33.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Jiaming Li
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	iodoacetamide derivatized residue
Instrument	Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2020-01-31 01:46:02	ID requested
1	2020-04-05 23:23:55	announced
⏵ 2	2020-04-30 03:48:54	announced	2020-04-30: Updated publication reference for PubMed record(s): 32243163.

Li J, Van Vranken JG, Paulo JA, Huttlin EL, Gygi SP, Selection of Heating Temperatures Improves the Sensitivity of the Proteome Integral Solubility Alteration Assay. J Proteome Res, 19(5):2159-2166(2020) [pubmed]

submitter keyword: Proteome Integral Solubility Alteration (PISA) Assay, human cell line lysate, LC-MS/MS

Steven P Gygi
contact affiliation	Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA
contact email	Steven_Gygi@hms.harvard.edu
lab head
Jiaming Li
contact affiliation	Harvard Medical School
contact email	jiaming_li@hms.harvard.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD017346
     1. Label: PRIDE project
     2. Name: Selection of Heating Temperatures Improves the Sensitivity of Proteome Integral Solubility Alteration (PISA) Assay