| 1993 |
The yeast PRE2 gene (highly homologous to human RING10/PSMB8, 60% identity) encodes a proteasome beta-subunit whose loss abolishes chymotrypsin-like activity of the 20S proteasome and causes accumulation of ubiquitinated proteins, establishing that this subunit is the catalytic component responsible for chymotryptic activity. |
Genetic complementation of pre2 mutants, missense mutation identification, biochemical assay of proteasome chymotryptic activity, ubiquitinated protein accumulation assay |
The Journal of biological chemistry |
High |
8383129
|
| 1992 |
A yeast homolog of RING10 (PSMB8), PRG1, is essential for cell viability, indicating that the proteasome function of this subunit is required for basic cellular processes. |
Genomic disruption of PRG1 in yeast showing lethality |
Gene |
Medium |
1452031
|
| 1993 |
LMP7 (PSMB8) is synthesized as a proprotein (~30 kDa) and undergoes post-translational N-terminal cleavage to yield the mature 23 kDa subunit that is incorporated into the 20S proteasome complex. |
Pulse-chase experiments, Western blot, anti-proteasome and anti-LMP7 immunoprecipitation |
European journal of biochemistry |
High |
8365398 8458375
|
| 1993 |
LMP7 protein undergoes post-translational cleavage of its N-terminus (pro-sequence) before incorporation into the proteasome, reducing from predicted ~30 kDa to 23 kDa; the cleaved form is the active proteasome subunit. |
Western blot of anti-proteasome immunoprecipitates, pulse-chase analysis |
European journal of immunology |
High |
8458375
|
| 1994 |
LMP7 (PSMB8) transfection into lymphoblasts or HeLa cells increases the Vmax of 20S and 26S proteasomes for cleavage after hydrophobic and basic residues (chymotrypsin-like and trypsin-like activities) without affecting hydrolysis after acidic residues; the magnitude of activity change is proportional to the amount of LMP7 incorporated. |
Gene transfection, fluorogenic peptide substrate assays of isolated 20S and 26S proteasomes |
Proceedings of the National Academy of Sciences of the United States of America |
High |
7937744
|
| 1994 |
LMP7 (PSMB8) and LMP2 are assembled into 20S proteasomes via 13-16S precursor complexes; pro-proteins of both subunits are processed within these preproteasome intermediates, and only the processed forms are part of active 20S proteasomes. |
Sedimentation analysis, pulse-chase labeling, immunoblotting of precursor and mature proteasome fractions |
Journal of molecular biology |
High |
8120905
|
| 1995 |
Incorporation of LMP7 (with or without LMP2) into 20S proteasomes alters cleavage site preference and changes the quality of peptide products generated from a defined polypeptide substrate (CMV IE pp89 25-mer), independently of IFN-gamma stimulation; both subunits together induce a drastic increase in positive cooperativity (Hill coefficient) between proteasome subunits. |
Stable transfection of LMP subunits, in vitro digestion of defined polypeptide, HPLC and mass spectrometry of cleavage products, fluorogenic substrate kinetics |
European journal of immunology |
High |
7589133
|
| 1995 |
The IFN-gamma-inducible 11S regulator (PA28) does not preferentially activate LMP7-containing proteasomes, but binding of PA28 to any proteasome preparation markedly changes the quality and quantity of peptide products; LMP7 content determines the cleavage product profile independently of PA28. |
Transfection of LMP subunits, in vitro 25-mer digestion with and without purified PA28, HPLC and electrospray mass spectrometry |
The Journal of biological chemistry |
High |
7559557
|
| 1996 |
LMP7 (PSMB8) replaces the constitutive subunit X (epsilon/β5c) upon IFN-gamma treatment; overexpression of X reduces hydrolysis after hydrophobic and basic residues—effects opposite to LMP7 transfection—demonstrating that loss of X contributes to the increased chymotryptic/tryptic activities seen after IFN-gamma induction. |
Gene transfection of subunit X into HeLa cells, fluorogenic peptide substrate assays, immunoblot quantification of subunit stoichiometry |
The Journal of biological chemistry |
High |
8663318
|
| 1997 |
LMP7 (PSMB8) is covalently labeled by active-site-directed radiolabeled chloromethane and diazomethane inhibitors of chymotrypsin-like activity, establishing LMP7 as a catalytic subunit that contributes to both trypsin-like and chymotrypsin-like proteasomal activities. |
Radiolabeled active-site inhibitor labeling of purified 20S and 26S proteasomes, RP-HPLC, SDS-PAGE, 2D-PAGE, immunoblotting with subunit-specific antibodies |
The Journal of biological chemistry |
High |
9312091
|
| 2000 |
The human proteasome maturation protein POMP (Ump1 homolog) is found only in 16S precursor complexes and not in mature 20S proteasomes; LMP7 propeptide deletion reduces incorporation efficiency and causes accumulation of precursor complexes with elevated POMP, indicating the propeptide assists maturation but is not strictly required for incorporation. An active-site threonine mutation does not affect LMP7 incorporation. |
2D gel analysis, subcellular fractionation, Northern blot, mutagenesis of LMP7 propeptide and active-site Thr, immunoblotting of precursor fractions in T2 cells |
Journal of molecular biology |
High |
10926487
|
| 2000 |
Overexpression of LMP2, LMP7, and MECL-1 together (triple transfectants) in cells markedly enhances MHC class I-restricted presentation of the LCMV NP118 epitope; in vitro, immunoproteasomes generate higher amounts of 11- and 12-mer precursor fragments containing NP118 than constitutive proteasomes. PA28 overexpression does not produce a comparable enhancement. |
Triple transfection, CTL killing assay for antigen presentation, in vitro peptide digestion with HPLC analysis of products |
Journal of immunology |
High |
10878350
|
| 2004 |
HCV NS3 protein directly interacts with the pro-sequence region (amino acids 1-40) of LMP7 via its protease domain (confirmed by yeast two-hybrid and in vitro binding and co-IP); this interaction does not affect NS3 protease activity in vitro, but cells stably replicating HCV subgenomic replicon show markedly reduced LMP7 immunoproteasome peptidase activities. |
Yeast two-hybrid screen, in vitro binding assay, co-immunoprecipitation, domain mapping, fluorogenic peptidase activity assay in HCV replicon cell line |
The Biochemical journal |
High |
15303969
|
| 2005 |
IRF-1 is required for IFN-gamma-dependent LMP7 transcription; a tetracycline-inducible IRF-1 system induces LMP7 expression, a specific IRF-1-binding genomic region in the LMP7 locus was identified, and IRF-1 knockdown by siRNA as well as IRF-1-/- mice confirmed that IRF-1 is the master transcriptional regulator driving concerted immunoproteasome subunit expression. |
Tet-inducible IRF-1 expression, siRNA knockdown, IRF-1 knockout mice, genomic footprinting/reporter assay to map IRF-1 binding region |
FEBS letters |
High |
15907481
|
| 2010 |
LMP7 (PSMB8) deficiency in mice results in significantly attenuated dextran sodium sulfate-induced colitis due to reduced NF-κB signaling; this is mechanistically linked to reduced secretion of proinflammatory cytokines and chemokines, decreased neutrophil infiltration, and diminished Th1/Th17 expansion. |
lmp7-/- mouse model, DSS colitis induction, NF-κB pathway analysis, cytokine/chemokine measurement, histological analysis |
Gut |
High |
20581238
|
| 2010 |
A homozygous missense mutation (Thr75Met) in PSMB8 causes JMP syndrome; the mutation disrupts the tertiary structure of β5i and results in significantly reduced chymotrypsin-like proteolytic activity of immunoproteasomes in patient lymphoblasts compared to normal cells. |
Homozygosity mapping, direct sequencing, structural modeling, immunoproteasome chymotryptic activity assay in patient vs. normal lymphoblasts |
American journal of human genetics |
High |
21129723
|
| 2011 |
A G201V mutation in PSMB8 (β5i) in Nakajo-Nishimura syndrome disrupts the β-sheet structure near the catalytic threonine, prevents efficient incorporation of β5i into immunoproteasome precursors, reduces total proteasome activity, and causes accumulation of ubiquitinated and oxidized proteins; this leads to increased IL-6 and IP-10 secretion and elevated p38 phosphorylation. |
Structural analysis (mutation position modeling), immunoproteasome assembly assay, proteasome activity assay, ubiquitinated/oxidized protein accumulation assay, cytokine ELISA, phospho-p38 immunoblot in patient cells |
Proceedings of the National Academy of Sciences of the United States of America |
High |
21852578
|
| 2011 |
A G197V mutation in PSMB8 increases assembly intermediates of immunoproteasomes, decreases proteasome function, causes ubiquitin-coupled protein accumulation, and leads to increased IL-6 expression in patient skin and B cells; PSMB8 knockdown inhibits murine and human adipocyte differentiation in vitro, and siRNA injection against Psmb8 in mouse skin reduces adipocyte tissue volume. |
Exome sequencing, proteasome activity assay, ubiquitin accumulation immunoblot, siRNA knockdown of Psmb8 in mouse skin/adipocyte differentiation assay |
The Journal of clinical investigation |
High |
21881205
|
| 2011 |
β5i/LMP7 deficiency in mice leads to impaired immunoproteasome function, failure to clear poly-ubiquitinated protein aggregates in cytokine-stressed cardiomyocytes during CVB3 myocarditis, impaired NF-κB activation, and increased apoptotic cell death, resulting in exacerbated acute myocardial damage despite identical viral load. |
β5i/LMP7 knockout mice, CVB3 infection model, ubiquitinated aggregate detection, NF-κB activation assay, apoptosis quantification, adoptive T cell transfer |
PLoS pathogens |
High |
21909276
|
| 2012 |
LMP7 propeptide (proLMP7) shows significantly higher chaperone activity than the β5 propeptide (proβ5) in promoting proteasome maturation; proLMP7 promotes integration of both immunosubunits and mixed proteasomes, and increased efficiency of proteasome maturation by proLMP7 is required for optimal MHC class I surface expression. LMP7 induction also increases total proteasome abundance in infected tissue. |
LMP7-deficient mouse infection model, IFNγ stimulation experiments, subunit integration assays, proteasome maturation kinetics, MHC class I surface expression by flow cytometry |
PloS one |
High |
22768135
|
| 2012 |
LMP7 deficiency or selective inhibition suppresses Th17 and Th1 differentiation while promoting regulatory T cell (Treg) development; mechanistically, LMP7 inhibition blocks STAT3 phosphorylation in developing Th17 cells and enhances SMAD phosphorylation in Tregs, and reduces STAT1 phosphorylation in Th1 cells. |
LMP7-/- CD4+ T cells and ONX 0914-treated WT T cells under polarizing conditions, phospho-STAT3/SMAD/STAT1 immunoblot, DSS colitis and T cell transfer colitis models in vivo |
Journal of immunology |
High |
22984077
|
| 2014 |
X-ray crystal structures of murine constitutive and immunoproteasome 20S core particles informed rational design of β5i (LMP7)-selective inhibitors; structural differences in the S1 pocket between β5c and β5i were exploited to achieve cell-permeable inhibitors with high selectivity. |
Structure-based drug design using X-ray crystallography of 20S proteasome complexes, selectivity profiling across all proteasome subunits |
Journal of medicinal chemistry |
High |
25006746
|
| 2016 |
miR-451 directly targets LMP7 (PSMB8) 3′UTR to suppress LMP7 expression, thereby inhibiting NF-κB activity and reducing proinflammatory cytokine transcription in mesangial cells; in db/db diabetic mice, increasing miR-451 inhibited LMP7/NF-κB and attenuated glomerular injury. |
Deep sequencing, dual-luciferase reporter assay, Western blot, chromatin immunoprecipitation, in vivo miR-451 overexpression in db/db mice |
Molecular and cellular endocrinology |
High |
27264074
|
| 2018 |
Co-inhibition of both LMP7 and LMP2 (but not LMP7 alone) is required to impair MHC class I surface expression, suppress IL-6 secretion, block Th17 differentiation, and strongly ameliorate experimental colitis and EAE; prolonged ONX 0914 exposure inhibits both subunits, explaining its efficacy. |
Selective LMP7 inhibitor PRN1126 vs. ONX 0914 comparison, LMP2+LMP7 dual inhibitor combination, DSS colitis, EAE models, IL-6 ELISA, MHC class I flow cytometry |
EMBO reports |
High |
30279279
|
| 2019 |
β5i (PSMB8) interacts with ATG5, promoting its degradation, thereby inhibiting autophagy and driving pathological cardiac hypertrophy; β5i knockout attenuates hypertrophy, and ATG5 knockdown or autophagy inhibition reverses the β5i-KO protection. |
β5i KO and transgenic overexpression mice, Ang II-induced hypertrophy model, Co-IP (β5i–ATG5 interaction), ATG5 degradation assay, autophagic flux assays, genetic epistasis (ATG5 KD + β5i KO) |
Science advances |
High |
31086810
|
| 2019 |
β5i (PSMB8) promotes ATRAP (AT1R-associated protein) degradation in atrial tissue; β5i upregulation by Ang II leads to ATRAP degradation, resulting in AT1R-mediated NF-κB activation, increased NADPH oxidase activity, TGF-β1/Smad signaling, and altered ion channel expression (Kir2.1, CX43), thereby driving atrial fibrillation. |
β5i KO and AAV9-β5i overexpression mice, Ang II infusion AF model, ATRAP co-IP and degradation assay, ATRAP overexpression rescue, NF-κB and TGF-β pathway assays |
Hypertension |
High |
30571551
|
| 2019 |
β5i (PSMB8) promotes ATRAP degradation in the retina; β5i KO restores Ang II-induced downregulation of ATRAP and attenuates AT1R-mediated downstream signaling, thereby reducing hypertensive retinopathy, while adenoviral β5i overexpression aggravates the phenotype. |
β5i KO and Ad-β5i overexpression mice, Ang II infusion retinopathy model, ATRAP degradation assay, AT1R downstream signaling analysis, ATRAP overexpression rescue |
Molecular therapy |
High |
31636038
|
| 2019 |
β5i (PSMB8) promotes PTEN degradation in DOCA-salt hypertensive hearts; β5i KO attenuates cardiac remodeling by restoring PTEN levels, and PTEN blockade reverses the protective effects of β5i KO, placing PTEN as a key substrate whose stability is regulated by β5i-dependent proteasomal degradation. |
β5i KO mice and PR-957 pharmacological inhibition, DOCA-salt hypertension model, PTEN degradation assay, AKT/mTOR/TGF-β/NF-κB pathway analysis, VO-OHpic rescue experiment |
Journal of molecular and cellular cardiology |
High |
31629736
|
| 2020 |
β5i (PSMB8) promotes atherosclerosis by targeting MERTK for degradation via NF-κB-dependent suppression of Mertk transcription; β5i deletion reduces IκBα degradation, inhibits NF-κB, increases MERTK expression, and enhances efferocytosis of apoptotic cells in atherosclerotic lesions. |
β5i/Apoe double KO mice, bone marrow transplantation, ATD-fed mouse atherosclerosis model, efferocytosis assay, MERTK expression, IκBα/NF-κB pathway analysis, PR-957 pharmacological treatment |
The Journal of pathology |
High |
31758542
|
| 2020 |
β5i (PSMB8) promotes macrophage pyroptosis during abdominal aortic aneurysm formation via activation of NF-κB and upregulation of NLRP3; β5i inhibition or knockout decreases macrophage pyroptosis and AAA severity through the IκB/NF-κB pathway. |
β5i KO mice, PR-957 treatment, AAA mouse model, pyroptosis markers in tissue and BMDMs, OXLDL stimulation, NF-κB/NLRP3 pathway analysis |
Biochemical and biophysical research communications |
Medium |
33019975
|
| 2021 |
M3258, an orally bioavailable, reversible, highly selective LMP7 (β5i) inhibitor, was developed through structure-based optimization using X-ray crystal structures; it demonstrates potent suppression of LMP7 activity, ubiquitinated protein turnover, and induces apoptosis in multiple myeloma cells in vitro and in vivo. |
Structure-based drug design (X-ray crystallography), selectivity profiling, multiple myeloma xenograft models, ubiquitin turnover assay, apoptosis assay |
Journal of medicinal chemistry |
High |
34228444
|
| 2018 |
Lmp7 (PSMB8) levels and activity are significantly increased in a mouse model of α-synuclein aggregation and in post-mortem human PD and dementia with Lewy bodies brains; the immunoproteasome degrades α-synuclein aggregates and generates potentially antigenic peptides from them. |
Quantitative proteomics (6215 proteins), immunoproteasome activity assay in mouse model and human post-mortem tissue, α-synuclein aggregate degradation assay |
EBioMedicine |
Medium |
29759483
|
| 2017 |
LMP7 is required in radioresistant thymic cells (likely mTECs) to prevent excessive negative selection of GP118-125-specific T cell precursors; LMP7-deficient mice completely lack GP118-125-specific CD8+ T cells, which is restored by bone marrow chimeras showing LMP7 requirement in non-hematopoietic cells. |
LMP7 KO mice, LCMV infection model, bone marrow chimera generation, adoptive transfer of LMP7-deficient CD8+ T cells into RAG1-/- mice, tetramer staining |
European journal of immunology |
High |
29067678
|
| 2022 |
PSMB8 (β5i) expression is upregulated in manganese-exposed microglia via PERK signaling; selective PSMB8 inhibition reduces neuroinflammation (TNF-α, iNOS, CCL12 production, microglial activation) and inhibition of PSMB8 reduces NF-κB p65 phosphorylation, placing PSMB8 in a PERK/NF-κB signaling axis during Mn neurotoxicity. |
In vivo Mn exposure mouse model, BV2 cell culture, selective PSMB8 inhibitor treatment, PERK inhibitor, NF-κB pathway analysis, learning/memory tests, Golgi staining |
Food and chemical toxicology |
Medium |
35378207
|
| 2018 |
Apelin inhibits epithelial-mesenchymal transition (EMT) of podocytes in diabetic mice through downregulation of β5i expression; β5i KO mice confirmed the dependence of apelin's anti-EMT effect on β5i; mechanistically, apelin decreases β5i expression, which reduces degradation of pIκB, promotes IκB nuclear translocation, inhibits NF-κB-driven TGF-β expression, and decreases Smad pathway activation. |
β5i KO mice, diabetic mouse model, apelin treatment, pIκB degradation assay, IκB localization, TGF-β/Smad pathway analysis |
Cell death & disease |
Medium |
30301930
|