The anti-inflammatory drug BAY 11-7082 suppresses the MyD88-dependent signalling network by targeting the ubiquitin system

Autor: Axel Knebel, Lorna Plater, Christoph H. Emmerich, Sam Strickson, Natalia Shpiro, Philip Cohen, David G. Campbell, Maria Stella Ritorto
Rok vydání: 2013
Předmět:
Lipopolysaccharides
Ubiquitin-conjugating enzyme
Biochemistry
DMEM
Dulbecco’s modified Eagle’s medium

Mice
linear ubiquitin assembly complex (LUBAC)
K63-pUb
Lys63-linked polyubiquitin

NF-KappaB Inhibitor alpha
Ubiquitin
MS/MS
tandem MS

nuclear factor κB (NF-κB)
pUb
polyubiquitin

TRAF
tumour-necrosis-factor-receptor-associated factor

Sulfones
RBR
RING-between-RING
TAB
TAK1-binding protein

IKK
IκB kinase

IκB
inhibitor of NF-κB

GFP
green fluorescent protein

HOIP
haem-oxidized IRP2 ligase-1-interacting protein

NEMO
NF-κB essential modifier

K48-pUb
Lys48-linked polyubiquitin

biology
LUBAC
linear ubiquitin assembly complex

Kinase
NEDD8
neural-precursor-cell-expressed developmentally down-regulated 8

Ubiquitin ligase
HRMS
high-resolution mass spectra

GAPDH
glyceraldehyde-3-phosphate dehydrogenase

TBK1
tumour-necrosis-factor-receptor-associated factor-associated NF-κB activator-binding kinase 1

JNK
c-Jun N-terminal kinase

LPS
lipopolysaccharide

Phosphorylation
I-kappa B Proteins
ubiquitin conjugating 13 (Ubc13)
NF-κB
nuclear factor κB

DLBCL
diffuse large B-cell lymphoma

PAMP
pathogen-associated molecular pattern

Signal Transduction
Research Article
Proteasome Endopeptidase Complex
Molecular Sequence Data
lymphoma
MALDI–TOF
matrix-assisted laser-desorption ionization–time-of-flight

ERK
extracellular-signal-regulated kinase

HEK
human embryonic kidney

Ubc
ubiquitin conjugating

Cell Line
Tumor

Nitriles
MyD88
myeloid differentiation factor 88

Animals
Humans
Amino Acid Sequence
Molecular Biology
HIF1α
hypoxia-inducible factor 1α

IL-1R
IL-1 receptor

HTLV-1
human T-cell lymphotropic virus 1

Macrophages
HEK 293 cells
Ubiquitination
NF-kappa B p50 Subunit
Receptors
Interleukin-1

myeloid differentiation factor 88 (MyD88)
TAK1
transforming growth factor β-activated kinase 1

Cell Biology
Hypoxia-Inducible Factor 1
alpha Subunit

Molecular biology
IL
interleukin

UBE
ubiquitin-activating enzyme

IκBα
proteasome
Gene Expression Regulation
Proteasome
IRAK
IL-receptor-associated kinase

Myeloid Differentiation Factor 88
Ubiquitin-Conjugating Enzymes
biology.protein
MAPK
mitogen-activated protein kinase

Interleukin-1
DAPI
4′
6-diamidino-2-phenylindole
Zdroj: Biochemical Journal
ISSN: 1470-8728
0264-6021
DOI: 10.1042/bj20121651
Popis: The compound BAY 11-7082 inhibits IκBα [inhibitor of NF-κB (nuclear factor κB)α] phosphorylation in cells and has been used to implicate the canonical IKKs (IκB kinases) and NF-κB in >350 publications. In the present study we report that BAY 11-7082 does not inhibit the IKKs, but suppresses their activation in LPS (lipopolysaccharide)-stimulated RAW macrophages and IL (interleukin)-1-stimulated IL-1R (IL-1 receptor) HEK (human embryonic kidney)-293 cells. BAY 11-7082 exerts these effects by inactivating the E2-conjugating enzymes Ubc (ubiquitin conjugating) 13 and UbcH7 and the E3 ligase LUBAC (linear ubiquitin assembly complex), thereby preventing the formation of Lys63-linked and linear polyubiquitin chains. BAY 11-7082 prevents ubiquitin conjugation to Ubc13 and UbcH7 by forming a covalent adduct with their reactive cysteine residues via Michael addition at the C3 atom of BAY 11-7082, followed by the release of 4-methylbenzene-sulfinic acid. BAY 11-7082 stimulated Lys48-linked polyubiquitin chain formation in cells and protected HIF1α (hypoxia-inducible factor 1α) from proteasomal degradation, suggesting that it inhibits the proteasome. The results of the present study indicate that the anti-inflammatory effects of BAY 11-7082, its ability to induce B-cell lymphoma and leukaemic T-cell death and to prevent the recruitment of proteins to sites of DNA damage are exerted via inhibition of components of the ubiquitin system and not by inhibiting NF-κB.
Databáze: OpenAIRE
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