Dataset Information

System-wide identification of enzyme substrates by thermal analysis (SIESTA)

ABSTRACT: Despite the immense importance of enzyme-substrate reactions, there is a lack of general and unbiased tools for identifying the molecular components participating in these reactions on a cellular level. Here we developed a universal method called System-wide Identification of Enzyme Substrates by Thermal Analysis (SIESTA). The approach assumes that post-translational modification of the substrate protein changes its thermal stability, and applies the concept of specificity to reveal the potential substrates. For selenoprotein thioredoxin reductase 1, SIESTA confirmed several known protein substrates and suggested novel candidates. For poly-(ADP-ribose) polymerase-10, SIESTA revealed a number of PARP10 putative substrates, which were confirmed by targeted mass spectrometry and functional assays. Wider application of SIESTA can enhance our understanding of the role of enzymes in homeostasis and disease, and facilitate drug discovery.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Colon Epithelial Cell

DISEASE(S): Colon Cancer

SUBMITTER: Amir Ata Saei

LAB HEAD: Roman Zubarev

PROVIDER: PXD010554 | Pride | 2021-01-11

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	PARP10_substrate_C_Rep1_Frac1.raw	Raw
	PARP10_substrate_C_Rep1_Frac2.raw	Raw
	PARP10_substrate_C_Rep1_Frac3.raw	Raw
	PARP10_substrate_C_Rep1_Frac4.raw	Raw
	PARP10_substrate_C_Rep1_Frac5.raw	Raw

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Publications

System-wide identification and prioritization of enzyme substrates by thermal analysis.

Saei Amir Ata AA Beusch Christian M CM Sabatier Pierre P Wells Juan Astorga JA Gharibi Hassan H Meng Zhaowei Z Chernobrovkin Alexey A Rodin Sergey S Näreoja Katja K Thorsell Ann-Gerd AG Karlberg Tobias T Cheng Qing Q Lundström Susanna L SL Gaetani Massimiliano M Végvári Ákos Á Arnér Elias S J ESJ Schüler Herwig H Zubarev Roman A RA

Nature communications 20210226 1

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 the ...[more]

PMID: 33637753

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