Thermal Proteome Profiling for Drug Target Identification and Probing of Protein States

Sauer, Patricia; Bantscheff, Marcus

doi:10.1007/978-1-0716-3457-8_5

Cited by 6 publications

(3 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In cellular thermal shift assays (CETSAs) [117], cells are incubated with the compounds to be tested, then lysed, and then the lysates are incubated for a given time period in a thermal cycler at various temperatures, starting at 37 °C [118]. Precipitated proteins are removed by centrifugation, and the supernatants are subjected to suitable functional assays [118], to oneor two-dimensional gel electrophoresis [119], or to LC-MS/MS [120,121]. Classical CETSA-MS is performed by incubating cells or lysates with the compounds to be tested or with a solvent control at ten different temperatures, followed by centrifugation of the precipitates, trypsinization and labeling of the soluble proteins with commercially available TMTs [122], or alternative labeling agents [123] and LC-MS analysis of the ten combined compound and control samples [124].…”

Section: Protein Stability-based Methodsmentioning

confidence: 99%

Investigating Antiprotozoal Chemotherapies with Novel Proteomic Tools—Chances and Limitations: A Critical Review

Müller,

Boubaker,

Müller

et al. 2024

IJMS

View full text Add to dashboard Cite

Identification of drug targets and biochemical investigations on mechanisms of action are major issues in modern drug development. The present article is a critical review of the classical “one drug”—“one target” paradigm. In fact, novel methods for target deconvolution and for investigation of resistant strains based on protein mass spectrometry have shown that multiple gene products and adaptation mechanisms are involved in the responses of pathogens to xenobiotics rather than one single gene or gene product. Resistance to drugs may be linked to differential expression of other proteins than those interacting with the drug in protein binding studies and result in complex cell physiological adaptation. Consequently, the unraveling of mechanisms of action needs approaches beyond proteomics. This review is focused on protozoan pathogens. The conclusions can, however, be extended to chemotherapies against other pathogens or cancer.

show abstract

Section: Protein Stability-based Methodsmentioning

confidence: 99%

Investigating Antiprotozoal Chemotherapies with Novel Proteomic Tools—Chances and Limitations: A Critical Review

Müller,

Boubaker,

Müller

et al. 2024

IJMS

View full text Add to dashboard Cite

show abstract

“…In cellular thermal shift assays (CETSA) [109], cells are incubated with the compounds to be tested, lysed and the lysates are incubated for a given time period in a thermal cycler at various temperatures starting at 37°C [110]. Precipitated proteins are removed by centrifugation, and the supernatants are subjected to suitable functional assays [110], to one or two-dimensional gel electrophoresis [111] or to LC-MS/MS [112,113]. Classical CETSA-MS is performed by incubating cells or lysates with the compounds to be tested or with a solvent control at ten different temperatures followed by centrifugation of the precipitates, trypsinization and labelling of the soluble proteins with commercially available TMTs [114] or alternative labelling agents [115] and LC-MS analysis of the ten combined compound and control samples [116].…”

Section: Thermal Proteome Profilingmentioning

confidence: 99%

Investigating Antiprotozoal Chemotherapies with Novel Proteomic Tools – Chances and Limitations. A Critical Review

Müller,

Boubaker,

Müller

et al. 2024

Preprint

View full text Add to dashboard Cite

Identification of drug targets and biochemical investigations on mechanisms of action is a are major issues in modern drug development. The present article reviews the classical “one drug” – “one target” paradigm. Novel methods for target deconvolution and for investigation of resistant strains based on protein mass spectrometry have shown that multiple gene products and adapta-tion mechanisms are involved in responses of pathogens to xenobiotics. This review is focused on protozoan pathogens. The conclusions can, however, be extended to chemotherapies against other pathogens or cancer. For this review we have searched Pubmed (pubmed.ncbi.nlm.nih.gov) and Web of Science (webofscience.com) using the key words listed below (status May 2024)

show abstract

“…On the other hand, methods creating high amounts of data, like mass spectrometry-based thermal proteome profiling (TPP), have gained interest, which gives insight at a proteomewide level. TTP-type methods are like CESTA and can provide information regarding the state of the protein and interactions, which is used to recognize the off-target effects of the drugs [19][20][21]. Another practical example of CESTA is to determine the stability of the membrane protein without detergents.…”

Section: Introductionmentioning

confidence: 99%

Fluorescence-Based Protein Stability Monitoring—A Review

Gooran,

Kopra

2024

IJMS

View full text Add to dashboard Cite

Proteins are large biomolecules with a specific structure that is composed of one or more long amino acid chains. Correct protein structures are directly linked to their correct function, and many environmental factors can have either positive or negative effects on this structure. Thus, there is a clear need for methods enabling the study of proteins, their correct folding, and components affecting protein stability. There is a significant number of label-free methods to study protein stability. In this review, we provide a general overview of these methods, but the main focus is on fluorescence-based low-instrument and -expertise-demand techniques. Different aspects related to thermal shift assays (TSAs), also called differential scanning fluorimetry (DSF) or ThermoFluor, are introduced and compared to isothermal chemical denaturation (ICD). Finally, we discuss the challenges and comparative aspects related to these methods, as well as future opportunities and assay development directions.

show abstract

Thermal Proteome Profiling for Drug Target Identification and Probing of Protein States

Cited by 6 publications

References 25 publications

Investigating Antiprotozoal Chemotherapies with Novel Proteomic Tools—Chances and Limitations: A Critical Review

Investigating Antiprotozoal Chemotherapies with Novel Proteomic Tools—Chances and Limitations: A Critical Review

Investigating Antiprotozoal Chemotherapies with Novel Proteomic Tools – Chances and Limitations. A Critical Review

Fluorescence-Based Protein Stability Monitoring—A Review

Contact Info

Product

Resources

About