Infection with <i>Salmonella typhimurium</i> has no effect on the composition and cleavage specificity of the 20S proteasome in human lymphoid cells

Marcilla, Miguel; Castro, José A. Łópez de; Castaño, José G.; Álvarez, Iñaki

doi:10.1111/j.1365-2567.2007.02624.x

Cited by 4 publications

(3 citation statements)

References 37 publications

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“…Western blot experiments were performed as described. 38 Samples of 20 µg of protein extracts were subjected to SDS-PAGE on 12% acrylamide gels in Trisglycine-SDS buffer. Electrophoretically separated proteins were subjected to semidry electrophoretic transfer onto nitrocellulose membranes at 0.8 mA/cm 2 for 1 h. Membranes were blocked for 30 min in T-PBS (PBS with 0.1% Tween 20) with 5% skimmed milk, and then incubated with the corresponding antibodies for 2 h at R.T. Membranes were washed three times in T-PBS and incubated with a HRP-labeled goat anti-mouse IgG antibody at a 1:5000 dilution for 1 h. Specific proteins were detected by ECL (Biological Industries Israel Beit Haemek Ltd., Ashrat, Israel).…”

Section: Methodssupporting

confidence: 71%

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Increased Apoptosis after Autoimmune Regulator Expression in Epithelial Cells Revealed by a Combined Quantitative Proteomics Approach

Colomé

Collado²,

Bech-Serra³

et al. 2010

J. Proteome Res.

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Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) is a rare autosomal recessive autoimmune disease, affecting many endocrine tissues. APECED is associated to the lack of function of a single gene called AutoImmune REgulator (AIRE). Aire knockout mice develop various autoimmune disorders affecting different organs, indicating that Aire is a key gene in the control of organ-specific autoimmune diseases. AIRE is mainly expressed by medullary thymic epithelial cells (mTECs), and its absence results in the loss of tolerance against tissue restricted antigens (TRAs). Aire induces the transcription of genes encoding for TRAs in mTECs. In this report, the analysis of AIRE's effect on the cellular proteome was approached by the combination of two quantitative proteomics techniques, 2D-DIGE and ICPL, using an AIRE-transfected and nontransfected epithelial cell line. The results showed increased levels of several chaperones, (HSC70, HSP27 and tubulin-specific chaperone A) in AIRE-expressing cells, while various cytoskeleton interacting proteins, that is, transgelin, caldesmon, tropomyosin alpha-1 chain, myosin regulatory light polypeptide 9, and myosin-9, were decreased. Furthermore, some apoptosis-related proteins were differentially expressed. Data were confirmed by Western blot and flow cytometry analysis. Apoptosis assays with annexin V and etoposide demonstrated that AIRE-positive cells suffer more spontaneous apoptosis and are less resistant to apoptosis induction.

show abstract

Section: Methodssupporting

confidence: 71%

“…Western blot experiments were performed as described . Samples of 20 μg of protein extracts were subjected to SDS-PAGE on 12% acrylamide gels in Tris-glycine-SDS buffer.…”

Section: Methodsmentioning

confidence: 99%

Increased Apoptosis after Autoimmune Regulator Expression in Epithelial Cells Revealed by a Combined Quantitative Proteomics Approach

Colomé

Collado²,

Bech-Serra³

et al. 2010

J. Proteome Res.

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“…Search terms were "proteasome", "proteasomal", "cleavage", "digestion", "immunoproteasome", "20S", "i20S", both alone and in various combinations. Ultimately, we identified 36 studies with relevant data (Table 1), from which we manually extracted individual cleavage sites, along with the parent peptide sequences from which they were derived 13,14,[23][24][25][26][27][28][29][30][31][32]15,[33][34][35][36][37][38][39][40][41][42]16,[43][44][45][46][47][17][18][19][20][21][22] . Proteasome types present in the observed system (constitutive, immunoproteasome, or mixed) were also annotated for each cleavage experiment.…”

Section: Methods: Collection and Processing Of In Vitro Digestion Map Data For Training And Testingmentioning

confidence: 99%

Pepsickle rapidly and accurately predicts proteasomal cleavage sites for improved neoantigen identification

Weeder

Wood

et al. 2021

Preprint

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Proteasomal cleavage is a key component in protein turnover, as well as antigen presentation and subsequent immune response. Herein we present pepsickle, an open-source tool for proteasomal cleavage prediction with better in vivo prediction performance (AUC) and computational speed than current models available in the field, and with the ability to predict sites based on both constitutive and immunoproteasome profiles. Post-hoc filtering of predicted patient neoepitopes using pepsickle significantly enriches for immune-responsive epitopes and may represent a significant opportunity to improve current epitope prediction and vaccine development pipelines.

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Trypanosoma cruzi: Effect of the infection on the 20S proteasome in non-immune cells

Faria¹,

Lima²,

Sá³

2008

Experimental Parasitology

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Infection with Salmonella typhimurium has no effect on the composition and cleavage specificity of the 20S proteasome in human lymphoid cells

Abstract: Infection with Salmonella typhimurium has no effect on the composition and cleavage specificity of the 20S proteasome in human lymphoid cells

Cited by 4 publications

References 37 publications

Increased Apoptosis after Autoimmune Regulator Expression in Epithelial Cells Revealed by a Combined Quantitative Proteomics Approach

Increased Apoptosis after Autoimmune Regulator Expression in Epithelial Cells Revealed by a Combined Quantitative Proteomics Approach

Pepsickle rapidly and accurately predicts proteasomal cleavage sites for improved neoantigen identification

Trypanosoma cruzi: Effect of the infection on the 20S proteasome in non-immune cells

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