System-wide identification and prioritization of enzyme substrates by thermal analysis
| Autor: | Sergey Rodin, Christian M. Beusch, Katja Näreoja, Herwig Schüler, Pierre Sabatier, Hassan Gharibi, Elias S.J. Arnér, Amir Ata Saei, Alexey Chernobrovkin, Massimiliano Gaetani, Zhaowei Meng, Ann-Gerd Thorsell, Ákos Végvári, Qing Cheng, Susanna L. Lundström, Roman A. Zubarev, Tobias Karlberg, Juan Astorga Wells |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Proteomics
0301 basic medicine Thioredoxin Reductase 1 Science General Physics and Astronomy Computational biology Article General Biochemistry Genetics and Molecular Biology Substrate Specificity 03 medical and health sciences 0302 clinical medicine Oxidoreductase Proto-Oncogene Proteins Drug Discovery Humans SIESTA (computer program) Polymerase chemistry.chemical_classification Multidisciplinary Mass spectrometry biology Drug discovery Carcinoma Biochemistry and Molecular Biology Proteins Substrate (chemistry) General Chemistry HCT116 Cells Enzymes 030104 developmental biology Enzyme chemistry biology.protein Selenoprotein Poly(ADP-ribose) Polymerases Protein Processing Post-Translational Proto-Oncogene Proteins c-akt 030217 neurology & neurosurgery Biokemi och molekylärbiologi Post-translational modifications |
| Zdroj: | Nature Communications, Vol 12, Iss 1, Pp 1-13 (2021) Nature Communications |
| Popis: | Despite the immense importance of enzyme–substrate reactions, there is a lack of general and unbiased tools for identifying and prioritizing substrate proteins that are modified by the enzyme on the structural level. Here we describe a high-throughput unbiased proteomics method called System-wide Identification and prioritization of Enzyme Substrates by Thermal Analysis (SIESTA). The approach assumes that the enzymatic post-translational modification of substrate proteins is likely to change their thermal stability. In our proof-of-concept studies, SIESTA successfully identifies several known and novel substrate candidates for selenoprotein thioredoxin reductase 1, protein kinase B (AKT1) and poly-(ADP-ribose) polymerase-10 systems. Wider application of SIESTA can enhance our understanding of the role of enzymes in homeostasis and disease, opening opportunities to investigate the effect of post-translational modifications on signal transduction and facilitate drug discovery. The global identification of enzyme substrates is still challenging. Here, the authors develop a method based on proteome-wide thermal shift assays to discover enzyme substrates directly from cell lysates, identifying known and novel oxidoreductase, kinase and poly-(ADP-ribose) polymerase substrates. |
| Databáze: | OpenAIRE |
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