Proteomics

Dataset Information

0

Spike-in Proteome to enhance Data-Independent Acquisition for Thermal Proteome Profiling


ABSTRACT: Target deconvolution interactions is essential for elucidating the molecular mechanisms, therapeutic efficacy and off-target toxicity of small molecule drugs. Thermal proteome profiling (TPP) is a robust popular method for identifying drug-protein interactions. Nevertheless, classical implementation of TPP using isobaric labelling of peptides is tedious, time-consuming and costly. This prompt the adoption of label-free approach with data-independent acquisition (DIA), but with substantial compromise in protein coverage and precision. To address these shortcomings, we improvise a spike-in proteome strategy for DIA with TPP to counteract the reduction in protein quantity following sample heating. Protein coverage, data completeness and quantification precision are significantly improved as result. Additionally, a calibration algorithm was developed to correct for spike-in effects on fold changes. The integration of DIA-TPP with the Matrix-Augmented Pooling Strategy (MAPS) to increase experiment throughput demonstrates performance comparable to existing TMT-TPP-MAPS. With this spike-in proteome strategy, we also successfully identified the thermal stabilization of CA13 by dorzolamide hydrochloride, as well as GSTZ1 and TDP1 of opicapone that eluded detection without the spike-in proteome.

INSTRUMENT(S): Orbitrap Exploris 480

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: qiqi Wang  

LAB HEAD: Chris Soon Heng Tan

PROVIDER: PXD056816 | Pride | 2025-03-11

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
DIA_1.raw Raw
DIA_10.raw Raw
DIA_11.raw Raw
DIA_12.raw Raw
DIA_13.raw Raw
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