TEAD-targeting small molecules induce a cofactor switch to regulate the Hippo pathway DOI Creative Commons
Alissa D. Guarnaccia, Thijs J. Hagenbeek, Wendy Lee

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 17, 2024

SUMMARY TEAD proteins are the main transcriptional effectors of Hippo signaling pathway and a pharmacological target in oncology. Most TEAD-targeting small molecules act by disrupting interaction with oncogenic activators YAP TAZ. Here, we describe an alternative mechanism for lipid pocket binding molecules. We report that select sulfonamide-containing compounds promote repressor VGLL4 to induce molecule-mediated cofactor switch from VGLL4. Chemically induced VGLL4–TEAD complexes counteract activity at chromatin repress pro-growth gene networks, including genes involved cellular proliferation mechanosignaling. is required anti-proliferative response these compounds, genetic deletion causes resistance vitro vivo . Our data reveal category facilitate repressive open up new understandings curbing deregulation.

Language: Английский

What’s new in single-cell proteomics DOI
Thy Truong, Ryan Kelly

Current Opinion in Biotechnology, Journal Year: 2024, Volume and Issue: 86, P. 103077 - 103077

Published: Feb. 14, 2024

Language: Английский

Citations

13
Functionalizing tandem mass tags for streamlining click-based quantitative chemoproteomics DOI Creative Commons
Nikolas R. Burton, Keriann M. Backus

Communications Chemistry, Journal Year: 2024, Volume and Issue: 7(1)

Published: April 10, 2024

Abstract Mapping the ligandability or potential druggability of all proteins in human proteome is a central goal mass spectrometry-based covalent chemoproteomics. Achieving this ambitious objective requires high throughput and coverage sample preparation liquid chromatography-tandem spectrometry analysis for hundreds to thousands reactive compounds chemical probes. Conducting chemoproteomic screens at scale benefits from technical innovations that achieve increased throughput. Here we realize vision by establishing silane-based cleavable linkers isotopically-labeled proteomics-tandem tag (sCIP-TMT) proteomic platform, which distinguished early pooling increases sCIP-TMT pairs custom click-compatible sCIP capture reagent readily functionalized yield with commercially available TMT reagents. Synthesis benchmarking 10-plex set reveal substantial decrease time together accuracy quantification. By screening focused four cysteine-reactive electrophiles, demonstrate utility target hunting, identifying 789 total liganded cysteines. Distinguished its compatibility established enrichment quantification protocols, expect will translate wide range applications.

Language: Английский

Citations

4
ATF6 promotes colorectal cancer growth and stemness by regulating the Wnt pathway DOI Creative Commons
Jeffrey J. Rodvold, Matthew Grimmer, Karen Ruiz

et al.

Cancer Research Communications, Journal Year: 2024, Volume and Issue: 4(10), P. 2734 - 2755

Published: Sept. 26, 2024

ATF6 intervention reduces colorectal cancer cell and organoid viability by interrupting dysregulated Wnt signaling, identifying a novel facilitator potential therapeutic target in cancer.

Language: Английский

Citations

3
Analysis of Protein Cysteine Acylation Using a Modified Suspension Trap (Acyl-Trap) DOI
Michael T. Forrester, Jacob R. Egol, Aleksandra Tata

et al.

Journal of Proteome Research, Journal Year: 2024, Volume and Issue: 23(8), P. 3716 - 3725

Published: July 15, 2024

Proteins undergo reversible S-acylation via a thioester linkage in vivo. S-palmitoylation, modification by C16:0 fatty acid, is common that mediates critical protein–membrane and protein–protein interactions. The most widely used assays, including acyl-biotin exchange acyl resin-assisted capture, utilize blocking of free Cys thiols, hydroxylamine-dependent cleavage the subsequent labeling nascent thiol. These assays generally require >500 μg protein input material per sample numerous reagent removal washing steps, making them laborious ill-suited for high throughput low applications. To overcome these limitations, we devised "Acyl-Trap", suspension trap-based assay utilizes thiol-reactive quartz to enable buffer hydroxylamine-mediated S-acyl enrichment. We show method compatible with protein-level detection S-acylated proteins (e.g., H-Ras) as well site identification quantification using "on trap" isobaric LC-MS/MS from little 20 input. In mouse brain, Acyl-Trap identified 279 reported sites 1298 previously unreported putative sites. Also described are conditions long-term hydroxylamine storage, which streamline assay. More generally, serves proof-of-concept PTM-tailored traps suitable both traditional chemoproteomic workflows.

Language: Английский

Citations

2
Ligand discovery by activity-based protein profiling DOI
Micah J. Niphakis, Benjamin F. Cravatt

Cell chemical biology, Journal Year: 2024, Volume and Issue: 31(9), P. 1636 - 1651

Published: Sept. 1, 2024

Language: Английский

Citations

2
Analysis of Protein Cysteine Acylation Using a Modified Suspension Trap (Acyl-Trap) DOI Creative Commons
Michael T. Forrester, Jacob R. Egol, Aleksandra Tata

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: March 27, 2024

Proteins undergo reversible S -acylation via a thioester linkage in vivo. -palmitoylation, modification by C16:0 fatty acid, is common that mediates critical protein-membrane and protein-protein interactions. The most widely used assays, including acyl-biotin exchange acyl resin-assisted capture, utilize blocking of free Cys thiols, hydroxylamine-dependent cleavage the subsequent labeling nascent thiol. These assays generally require >500 micrograms protein input material per sample numerous reagent removal washing steps, making them laborious ill-suited for high throughput low applications. To overcome these limitations, we devised Acyl-Trap, suspension trap-based assay utilizes thiol-reactive quartz to enable buffer hydroxylamine-mediated -acyl enrichment. We show method compatible with protein-level detection -acylated proteins (e.g. H-Ras) as well site identification quantification using on-trap isobaric LC-MS/MS from little 20 input. In mouse brain, Acyl-Trap identified 279 reported sites 1298 previously unreported putative sites. Also described are conditions long-term hydroxylamine storage, which streamlines assay. More generally, serves proof-of-concept PTM-tailored traps suitable both traditional chemoproteomic workflows.

Language: Английский

Citations

1
CySP3-96 enables scalable, streamlined, and low-cost sample preparation for cysteine chemoproteomic applications DOI Creative Commons
Flowreen Shikwana,

Beeta S. Heydari,

Samuel Ofori

et al.

Molecular & Cellular Proteomics, Journal Year: 2024, Volume and Issue: unknown, P. 100898 - 100898

Published: Dec. 1, 2024

Language: Английский

Citations

1
Celebrating Women in Proteomics and Metabolomics DOI Creative Commons
Ileana M. Cristea, Claire E. Eyers

Journal of Proteome Research, Journal Year: 2024, Volume and Issue: 23(8), P. 2675 - 2679

Published: Aug. 2, 2024

Language: Английский

Citations

0
TEAD-targeting small molecules induce a cofactor switch to regulate the Hippo pathway DOI Creative Commons
Alissa D. Guarnaccia, Thijs J. Hagenbeek, Wendy Lee

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 17, 2024

SUMMARY TEAD proteins are the main transcriptional effectors of Hippo signaling pathway and a pharmacological target in oncology. Most TEAD-targeting small molecules act by disrupting interaction with oncogenic activators YAP TAZ. Here, we describe an alternative mechanism for lipid pocket binding molecules. We report that select sulfonamide-containing compounds promote repressor VGLL4 to induce molecule-mediated cofactor switch from VGLL4. Chemically induced VGLL4–TEAD complexes counteract activity at chromatin repress pro-growth gene networks, including genes involved cellular proliferation mechanosignaling. is required anti-proliferative response these compounds, genetic deletion causes resistance vitro vivo . Our data reveal category facilitate repressive open up new understandings curbing deregulation.

Language: Английский

Citations

0