Journal: Journal of dairy science

Article Title: SRM SHOWS SPECIFIC HOST RESPONSE TO MASTITIS

doi: 10.3168/jds.2017-14312

Figure Lengend Snippet: Target proteins, their corresponding two proteotypic peptides and transitions used in the SRM analysis. Q3 ions that passed the manual verification step and used in this study for quantification are highlighted in bold. Q3 ions are named according to the precursor charge state and fragment ion position in the y ion series; hence 2y6 refers to a y6 fragment ion derived from the doubly charged precursor ion.

Article Snippet: Protein name Protein abbreviation Peptide sequence Transitions Alpha-1 antitrypsin A1AT ADLSGITK 2y5;2y6;2y7 ;3y3;3y4 Alpha-1 antitrypsin A1AT LSISETYDLK 2y5;2y6;2y7;2y8;2y9 Alpha 2 macroglobulin A2M IQHHTLLASPVR 2y8;3y5;3y6;3y7;3y8 Alpha 2 macroglobulin A2M LPPNVVEESAR 2y6;2y7;2y8;2y9;3y5 Calgranulin B (S100A9) CALGB ELPNFLK 2y4;2y5;2y6 ;3y3;3y4 Calgranulin B (S100A9) CALGB LGHYDTLIQK 2y6;2y7;3y4;3y5;3y6 Calgranulin C (S100A12) CALGC DQPTIDK 2y4;2y5 ;2y6;3y3;3y4 Calgranulin C (S100A12) CALGC VGHFDTLNK 2y5;2y6;3y3;3y4;3y5 Cathepsin C CATC NVHGINFVTPVR 2y7;2y9;3y4;3y5 ;3y6 Cathepsin C CATC TGNTSENVNVNTAR 2y10;2y11;2y7;2y8;2y9 CD14 CD14 LGAAQVPAQLLVAVLR 2y11;2y8;3y6;3y7;3y8 CD14 CD14 VQPQSLDLSHNSLR 3y5;3y6;3y7;3y8;3y9 Haptoglobin HP NQLVEVEK 2y4;2y5;2y6;2y7 ;3y3 Haptoglobin HP VGYVSGWGR 2y5;2y6;2y7;2y8 ;3y3 Interleukin 8 IL-8 THSTPFHPK 2y5;2y7;3y3;3y4;3y5 Interleukin 8 IL-8 VVQVFVK 2y4;2y5;2y6 ;3y3;3y4 Galectin 1 LGALS1 DAGAWGAEQR 2y5;2y6 ;2y7;2y8;3y3 Galectin 1 LGALS1 LPDGYEFK 2y4;2y5;2y6;2y7;3y3 Galectin 3 LGALS3 GNDVAFHFNPR 2y5;2y7;3y4;3y5;3y6 Galectin 3 LGALS3 IQVLVEPDHFK 2y7;2y9;3y5;3y6;3y7 Lactotransferrin LTF GEADALNLDGGYIYTAGK 2y10;2y11;3y6;3y7;3y8 Lactotransferrin LTF YYGYTGAFR 2y6;2y7;2y8 ;3y3; 3y4 Serum amyloid A3 SAA3 ADQFANEWGR 2y5;2y6;2y7;2y8 ;2y9 Serum amyloid A3 SAA3 ETIQGITDPLFK 2y6;2y8;3y4;3y5;3y6 Vanin-1 (Pantetheinase) VAN1 DSAPNTLSDLTTQALR 2y10;2y8;2y9;3y5;3y6 Vanin-1 (Pantetheinase) VAN1 NLDLLEGAVTSASK 2y10;2y11;2y12;2y9 ;3y6 Open in a separate window Target proteins, their corresponding two proteotypic peptides and transitions used in the SRM analysis.

Techniques: Derivative Assay, Sequencing

Journal: Journal of internal medicine

Article Title: The haptoglobin beta subunit sequesters HMGB1 toxicity in sterile and infectious inflammation

doi: 10.1111/joim.12619

Figure Lengend Snippet: (A) Schematics of human haptoglobin. The Human haptoglobin gene has two alleles and gives rise to three phenotypes termed Hp1-1, Hp2-2, and Hp 2–1. All three phenotypes are comprised of α (α1 or α2) and β subunits linked by disulfide bonds [11].

Article Snippet: Cloning and expression of human haptoglobin β subunit The expression plasmid construct encoding human haptoglobin (accession number {"type":"entrez-nucleotide","attrs":{"text":"XM_042621","term_id":"14779266","term_text":"XM_042621"}} XM_042621 ) β subunit (245 amino acids corresponding to 162-406 residues of haptoglobin holo-protein) was made available by GeneCopoeia Inc., (Germantown, MD).

Techniques:

Journal: Journal of internal medicine

Article Title: The haptoglobin beta subunit sequesters HMGB1 toxicity in sterile and infectious inflammation

doi: 10.1111/joim.12619

Figure Lengend Snippet: (A). Upper: 15N-1H- Heteronuclear Single Quantum Coherence (HSQC) spectra for Δ30-HMGB1 in the presence or absence of haptoglobin β. HMGB1 residues (8 amino acids) with significant chemical shift perturbations due to haptoglobin β binding are labeled. Lower: enlarged 15N-HSQC spectra of Δ30-HMGB1 free (black) and in complex with haptoglobin β (red). The 8 residues showing chemical shift changes seen with disulfide Δ30-HMGB1 upon complex formation with haptoglobin β are circled. Non-overlapping black and red resonance spectra indicate a significant chemical shift; hence, an interaction between proteins via that amino acid.

Article Snippet: Cloning and expression of human haptoglobin β subunit The expression plasmid construct encoding human haptoglobin (accession number {"type":"entrez-nucleotide","attrs":{"text":"XM_042621","term_id":"14779266","term_text":"XM_042621"}} XM_042621 ) β subunit (245 amino acids corresponding to 162-406 residues of haptoglobin holo-protein) was made available by GeneCopoeia Inc., (Germantown, MD).

Techniques: Binding Assay, Labeling

Journal: Journal of internal medicine

Article Title: The haptoglobin beta subunit sequesters HMGB1 toxicity in sterile and infectious inflammation

doi: 10.1111/joim.12619

Figure Lengend Snippet: (A) RAW 264.7 cells in 96-well culture plates were stimulated with HMGB1 in combination with various amounts of recombinant haptoglobin β for 16 hours. TNF released in the supernatants were measured. The effective concentration of haptoglobin β that inhibited 50% of released TNF (EC50) was approximately at 10 μg/ml (n = 5 experiments). *: p<0.05 vs. HMGB1 alone.

Article Snippet: Cloning and expression of human haptoglobin β subunit The expression plasmid construct encoding human haptoglobin (accession number {"type":"entrez-nucleotide","attrs":{"text":"XM_042621","term_id":"14779266","term_text":"XM_042621"}} XM_042621 ) β subunit (245 amino acids corresponding to 162-406 residues of haptoglobin holo-protein) was made available by GeneCopoeia Inc., (Germantown, MD).

Techniques: Recombinant, Concentration Assay

Journal: Journal of internal medicine

Article Title: The haptoglobin beta subunit sequesters HMGB1 toxicity in sterile and infectious inflammation

doi: 10.1111/joim.12619

Figure Lengend Snippet: (A) Surface plasmon resonance assay (SPR) of HMGB1 and haptoglobin β complexes to CD163. Recombinant human CD163 was coated on the sensor chip, the complexes of HMGB1 and haptoglobin β (1:1 molar ratio) at concentrations of 0, 32, 65, 125 or 250 nM were flow over the chip and the binding to CD163 (response units) was recorded. The Kd of HMGB1 and haptoglobin β complexes to CD163 is approximately 70 nM. Data are representative of 3 separate experiments.

Article Snippet: Cloning and expression of human haptoglobin β subunit The expression plasmid construct encoding human haptoglobin (accession number {"type":"entrez-nucleotide","attrs":{"text":"XM_042621","term_id":"14779266","term_text":"XM_042621"}} XM_042621 ) β subunit (245 amino acids corresponding to 162-406 residues of haptoglobin holo-protein) was made available by GeneCopoeia Inc., (Germantown, MD).

Techniques: SPR Assay, Recombinant, Binding Assay

Journal: Journal of internal medicine

Article Title: The haptoglobin beta subunit sequesters HMGB1 toxicity in sterile and infectious inflammation

doi: 10.1111/joim.12619

Figure Lengend Snippet: (A) The uptake of HMGB1 and haptoglobin (or haptoglobin β) complexes was measured by flow cytometry analysis. CD163 shRNA lentiviral or vector alone (WT) transduced primary human macrophages were incubated with FITC-labeled HMGB1 (1 μg/ml) plus haptoglobin (3 μg/ml) or haptoglobin β (1μg/ml) for 30 minutes. Uptake of FITC-HMGB1 was performed by staining for CD163 PE and FITC on cells. Data are representative from 5 separate experiments.

Article Snippet: Cloning and expression of human haptoglobin β subunit The expression plasmid construct encoding human haptoglobin (accession number {"type":"entrez-nucleotide","attrs":{"text":"XM_042621","term_id":"14779266","term_text":"XM_042621"}} XM_042621 ) β subunit (245 amino acids corresponding to 162-406 residues of haptoglobin holo-protein) was made available by GeneCopoeia Inc., (Germantown, MD).

Techniques: Flow Cytometry, shRNA, Plasmid Preparation, Incubation, Labeling, Staining

Journal: Journal of internal medicine

Article Title: The haptoglobin beta subunit sequesters HMGB1 toxicity in sterile and infectious inflammation

doi: 10.1111/joim.12619

Figure Lengend Snippet: (A–C) Administration of haptoglobin β subunit confers protection against lethal sepsis in wild type (A) and haptoglobin−/− mice (C) partly through reducing systemic cytokine accumulation and increase IL-10 release (B).

Article Snippet: Cloning and expression of human haptoglobin β subunit The expression plasmid construct encoding human haptoglobin (accession number {"type":"entrez-nucleotide","attrs":{"text":"XM_042621","term_id":"14779266","term_text":"XM_042621"}} XM_042621 ) β subunit (245 amino acids corresponding to 162-406 residues of haptoglobin holo-protein) was made available by GeneCopoeia Inc., (Germantown, MD).

Techniques:

Journal: Journal of internal medicine

Article Title: The haptoglobin beta subunit sequesters HMGB1 toxicity in sterile and infectious inflammation

doi: 10.1111/joim.12619

Figure Lengend Snippet: (A) C57BL/6 mice (male, 8–12 weeks old) were fasted overnight and received intraperitoneal injection of acetaminophen (APAP, 400 mg/kg, in 200 μl volume) to induced liver toxicity. Mice were given injections of haptoglobin β (200 μg per mouse, IP) or vehicle (PBS) control once a day for 5 days starting at 2 hours post-acetaminophen injection, and survival was monitored for two weeks (n = 20 per group). *: P < 0.05 vs. vehicle control group.

Article Snippet: Cloning and expression of human haptoglobin β subunit The expression plasmid construct encoding human haptoglobin (accession number {"type":"entrez-nucleotide","attrs":{"text":"XM_042621","term_id":"14779266","term_text":"XM_042621"}} XM_042621 ) β subunit (245 amino acids corresponding to 162-406 residues of haptoglobin holo-protein) was made available by GeneCopoeia Inc., (Germantown, MD).

Techniques: Injection, Control

Journal: Journal of internal medicine

Article Title: The haptoglobin beta subunit sequesters HMGB1 toxicity in sterile and infectious inflammation

doi: 10.1111/joim.12619

Figure Lengend Snippet: During infection or injury, HMGB1, a critical mediator in the final common pathway of inflammation, is actively secreted or passively released outside of the cells. Haptoglobin binds HMGB1 and inhibits its toxicity; and haptoglobin β subunit alone is sufficient to recapitulate the effects of the full-length haptoglobin. HMGB1-haptoglobin β complexes act via CD163 and elicit an anti-inflammatory response by stimulating HO-1 expression and IL-10 release; and by inhibiting pro-inflammatory cytokine release. Thus, haptoglobin β acts as an endogenous inhibitor of HMGB1 via a novel CD163-dependent anti-inflammatory pathway.

Article Snippet: Cloning and expression of human haptoglobin β subunit The expression plasmid construct encoding human haptoglobin (accession number {"type":"entrez-nucleotide","attrs":{"text":"XM_042621","term_id":"14779266","term_text":"XM_042621"}} XM_042621 ) β subunit (245 amino acids corresponding to 162-406 residues of haptoglobin holo-protein) was made available by GeneCopoeia Inc., (Germantown, MD).

Techniques: Infection, Expressing