Pirin does not bind to p65 or regulate NFkappaB-dependent gene expression DOI Creative Commons

Melissa Meschkewitz,

Erika M. Lisabeth,

A. Denaly Cab-Gomez

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Дек. 4, 2024

Pirin is a non-heme iron binding protein with variety of proposed functions including serving as co-activator p65 NFκB and quercetinase activity. We report here, failure to confirm pirin's primary mechanism, Fe(III)-pirin p65. Analytical size exclusion chromatography (SEC) fluorescence polarization (FP) studies did not detect an interaction. also found no effects pirin on TNFα-activated p65-regulated gene transcription using mouse embryonic fibroblasts (MEFs) from knockout knockdown NIH3T3 fibroblast cell line. TNFα - activated response mRNA was neither increased nor decreased in cells loss compared wildtype cells. Furthermore, immunofluorescence showed primarily cytoplasmic localization, nuclear most previous studies. This confirmed by fractionation analysis. show colocalization the endoplasmic reticulum (ER) marker disulfide-isomerase (PDI) well cyotoplasmic labeling. activity biochemical assays demonstrated competitive inhibition inhibitor CCG-257081. Cellular quercetin levels exposed

Язык: Английский

Pirin does not bind to p65 or regulate NFkappaB-dependent gene expression DOI Creative Commons

Melissa Meschkewitz,

Erika M. Lisabeth,

A. Denaly Cab-Gomez

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Дек. 4, 2024

Pirin is a non-heme iron binding protein with variety of proposed functions including serving as co-activator p65 NFκB and quercetinase activity. We report here, failure to confirm pirin's primary mechanism, Fe(III)-pirin p65. Analytical size exclusion chromatography (SEC) fluorescence polarization (FP) studies did not detect an interaction. also found no effects pirin on TNFα-activated p65-regulated gene transcription using mouse embryonic fibroblasts (MEFs) from knockout knockdown NIH3T3 fibroblast cell line. TNFα - activated response mRNA was neither increased nor decreased in cells loss compared wildtype cells. Furthermore, immunofluorescence showed primarily cytoplasmic localization, nuclear most previous studies. This confirmed by fractionation analysis. show colocalization the endoplasmic reticulum (ER) marker disulfide-isomerase (PDI) well cyotoplasmic labeling. activity biochemical assays demonstrated competitive inhibition inhibitor CCG-257081. Cellular quercetin levels exposed

Язык: Английский

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