Improved deconvolution of natural products’ protein targets using diagnostic ions from chemical proteomics linkers DOI Creative Commons
Andreas Wiest, Pavel Kielkowski

Beilstein Journal of Organic Chemistry, Год журнала: 2024, Номер 20, С. 2323 - 2341

Опубликована: Сен. 12, 2024

Identification of interactions between proteins and natural products or similar active small molecules is crucial for understanding their mechanism action on a molecular level. To search elusive, often labile, low-abundant conjugates compounds, chemical proteomics introduces feasible strategy that allows to enrich detect these conjugates. Recent advances in mass spectrometry techniques algorithms provide unprecedented depth proteome coverage the possibility desired modified peptides with high sensitivity. The 'linker' connecting an compound-protein conjugate detection tag critical component all proteomic workflows. In this review, we discuss properties applications different linkers special focus fragmentation releasing diagnostic ions how may improve confidence identified compound-peptide application advanced options improves identification rates help identify otherwise difficult find compounds proteins, which result from unperturbed conditions, thus are physiological relevance.

Язык: Английский

Development and Characterization of a Fluorinated MS-Cleavable Cross-Linker for Structural Proteomics DOI Creative Commons
Sorokin Av, Frank Hause, Christian Ihling

и другие.

Journal of the American Society for Mass Spectrometry, Год журнала: 2025, Номер 36(6), С. 1410 - 1413

Опубликована: Июнь 4, 2025

Cross-linking mass spectrometry (XL-MS) is an important method for studying three-dimensional protein structures and mapping protein-protein interactions. Some limitations of XL-MS still consist its use in-cell in vivo applications. To date, cross-linking reagents are urgently needed that can efficiently penetrate the cell membrane to comprehensively map interaction networks intact cells. In this study, fluorinated MS-cleavable cross-linker bis(pentafluorophenyl) ureido-4,4'-dibutyrate (DPFU) described. DPFU based on disuccinimidyl dibutyric urea (DSBU) with aim balancing hydrophobicity solubility improve permeability. was evaluated behavior different detergent solutions optimize conditions potential application live Using bovine serum albumin (BSA) as a model protein, experiments were conducted across temperatures concentrations. Solubility assays identified sodium dodecyl sulfate (SDS) effective enhancing aqueous environment. yielded fewer cross-links BSA than DSBU, highlighting regarding efficiency under similar experimental conditions. This study provides first insights into cross-linkers, suggesting further optimization broader future studies.

Язык: Английский

Процитировано

0

Improved deconvolution of natural products’ protein targets using diagnostic ions from chemical proteomics linkers DOI Creative Commons
Andreas Wiest, Pavel Kielkowski

Beilstein Journal of Organic Chemistry, Год журнала: 2024, Номер 20, С. 2323 - 2341

Опубликована: Сен. 12, 2024

Identification of interactions between proteins and natural products or similar active small molecules is crucial for understanding their mechanism action on a molecular level. To search elusive, often labile, low-abundant conjugates compounds, chemical proteomics introduces feasible strategy that allows to enrich detect these conjugates. Recent advances in mass spectrometry techniques algorithms provide unprecedented depth proteome coverage the possibility desired modified peptides with high sensitivity. The 'linker' connecting an compound-protein conjugate detection tag critical component all proteomic workflows. In this review, we discuss properties applications different linkers special focus fragmentation releasing diagnostic ions how may improve confidence identified compound-peptide application advanced options improves identification rates help identify otherwise difficult find compounds proteins, which result from unperturbed conditions, thus are physiological relevance.

Язык: Английский

Процитировано

0