ArticleJournal of mass spectrometry : JMS2025
Top-Down Thermal Proteome Profiling (TD-TPP) for Functional Characterization of the Intact Proteoforms in Complex Samples.
Article in Journal of mass spectrometry : JMS, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
0 citing papers in PubMed.
No citing paper in PubMed yet.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
4 authors.
Funding
Abstract
Protein function can vary due to changes in primary structure, such as post-translational modification (PTM), truncation, or amino acid replacement, and functional proteomics methods focus on elucidating changes in the function of proteins in biological pathways. Thermal proteome profiling (TPP) is a powerful functional proteomics approach that analyzes the thermal stability of proteins by exposing them to a temperature gradient to facilitate thermal denaturation, followed by analysis of the remaining folded proteins. Current TPP methods, however, utilize bottom-up methods that require protein digestion and can obscure relevant information regarding the structure of the intact proteoform. In this study, we have developed a top-down (TD) TPP approach to study intact proteoform stability as well as a high-throughput data analysis pipeline for label-free quantitative analysis and identification. We benchmarked this platform using two proteoforms of standard proteins, β-lactoglobulin A and β-lactoglobulin B (βLG-A and βLG-B), and found that βLG-A is slightly stabilized compared with βLG-B as a result of two amino acid substitutions. Additionally, we utilized this platform to detect protein thermal stability shifts induced by ligand binding, using carbonic anhydrase and its known inhibitor, acetazolamide. Our results demonstrated that the TD-TPP platform effectively detected the stabilization of a standard protein upon ligand binding. Furthermore, we adapted the TD-TPP platform for high-throughput thermal stability profiling of the intact E. coli proteome, enabling the characterization of intact proteoform-level thermal stability in complex biological samples. We performed thermal profiling of 72 identified proteoforms and 91 proteoform features (e.g., 163 total proteoforms) from intact E. coli lysate, and found that the melting points of these proteoforms correlated well with melting points determined using bottom-up TPP methods. Overall, the TD-TPP platform is capable of profiling thermal stability for standard proteins and intact proteoforms in complex biological samples.
Indexed as
Identifiers
What OpenQuestion holds
Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.