Characterization of Native COMPASS Complex in Urothelial Carcinoma Cells by Size Exclusion Chromatography.

Christoph Peter, Wolfgang A Schulz, Patcharawalai Whongsiri

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Article in Methods in molecular biology (Clifton, N.J.), 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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5 · Who and what money

Authors and funding

3 authors at 2 institutions in 2 countries.

Christoph PeterInstitute of Molecular Medicine I, Medical Faculty, Heinrich Heine University Düsseldorf, Düsseldorf, Germany.

Wolfgang A SchulzDepartment of Urology, Medical Faculty and University Hospital Düsseldorf, Heinrich-Heine-University, Düsseldorf, Germany.

Patcharawalai WhongsiriDepartment of Biochemistry and Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand. patcharawalai.wh@chula.ac.th.

Heinrich Heine University Düsseldorf · DEChulalongkorn University · TH

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

The human COMPASS complexes regulate gene expression during development and cell differentiation. Three distinct subunits, KMT2C, KMT2D, and KDM6A (also known as UTX), are frequently mutated in urothelial carcinoma, possibly disrupting the formation of functional COMPASS complexes. Here, we describe methods to evaluate the formation of these large native protein complexes in urothelial carcinoma (UC) cell lines harboring different mutations in KMT2C/D. To this end COMPASS complexes were purified from nuclear extracts by size exclusion chromatography (SEC) using a Sepharose 6 column. SEC fractions were then separated by 3-8% Tris-acetate gradient polyacrylamide gel and the COMPASS complex subunits KMT2C, UTX, WDR5, and RBBP5 were detected by immunoblotting. In this fashion, the formation of a COMPASS complex could be observed in UC cells with wild-type but not in cells with mutant KMT2C and KMTD.

Indexed as

Carcinoma, Transitional CellUrinary Bladder NeoplasmsCell DifferentiationCell NucleusChromatography, GelHumansIntracellular Signaling Peptides and ProteinsIntracellular Signaling Peptides and ProteinsWDR5 protein, humanCOMPASS complexGradient SDS–PAGEKMT2Nuclear protein fractionationSize exclusion chromatographyUrothelial cancerUTX

Identifiers

PMID37410229

DOI10.1007/978-1-61779-821-4_17

OpenAlexW4383302488

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.