Distinct Proteasome Subpopulations in the Alveolar Space of Patients with the Acute Respiratory Distress Syndrome
There is increasing evidence that proteasomes have a biological role in the extracellular alveolar space, but inflammation could change their composition. We tested whether immunoproteasome protein-containing subpopulations are present in the alveolar space of patients with lung inflammation evoking...
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Wiley
2012-01-01
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| Series: | Mediators of Inflammation |
| Online Access: | http://dx.doi.org/10.1155/2012/204250 |
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| author | S. U. Sixt R. Alami J. Hakenbeck M. Adamzik A. Kloß U. Costabel P. R. Jungblut B. Dahlmann J. Peters |
| author_facet | S. U. Sixt R. Alami J. Hakenbeck M. Adamzik A. Kloß U. Costabel P. R. Jungblut B. Dahlmann J. Peters |
| author_sort | S. U. Sixt |
| collection | DOAJ |
| description | There is increasing evidence that proteasomes have a biological role in the extracellular alveolar space, but inflammation could change their composition. We tested whether immunoproteasome protein-containing subpopulations are present in the alveolar space of patients with lung inflammation evoking the acute respiratory distress syndrome (ARDS). Bronchoalveolar lavage (BAL) supernatants and cell pellet lysate from ARDS patients (n=28) and healthy subjects (n=10) were analyzed for the presence of immunoproteasome proteins (LMP2 and LMP7) and proteasome subtypes by western blot, chromatographic purification, and 2D-dimensional gelelectrophoresis. In all ARDS patients but not in healthy subjects LMP7 and LMP2 were observed in BAL supernatants. Proteasomes purified from pooled ARDS BAL supernatant showed an altered enzyme activity ratio. Chromatography revealed a distinct pattern with 7 proteasome subtype peaks in BAL supernatant of ARDS patients that differed from healthy subjects. Total proteasome concentration in BAL supernatant was increased in ARDS (971 ng/mL ± 1116 versus 59±25; P<0.001), and all fluorogenic substrates were hydrolyzed, albeit to a lesser extent, with inhibition by epoxomicin (P=0.0001). Thus, we identified for the first time immunoproteasome proteins and a distinct proteasomal subtype pattern in the alveolar space of ARDS patients, presumably in response to inflammation. |
| format | Article |
| id | doaj-art-77ac9f1259a54ff79e01edf988d8596d |
| institution | OA Journals |
| issn | 0962-9351 1466-1861 |
| language | English |
| publishDate | 2012-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Mediators of Inflammation |
| spelling | doaj-art-77ac9f1259a54ff79e01edf988d8596d2025-08-20T02:07:09ZengWileyMediators of Inflammation0962-93511466-18612012-01-01201210.1155/2012/204250204250Distinct Proteasome Subpopulations in the Alveolar Space of Patients with the Acute Respiratory Distress SyndromeS. U. Sixt0R. Alami1J. Hakenbeck2M. Adamzik3A. Kloß4U. Costabel5P. R. Jungblut6B. Dahlmann7J. Peters8Klinik für Anästhesiologie und Intensivmedizin, Universität Duisburg-Essen, Universitätsklinikum Essen, 45122 Essen, GermanyKlinik für Anästhesiologie und Intensivmedizin, Universität Duisburg-Essen, Universitätsklinikum Essen, 45122 Essen, GermanyKlinik für Anästhesiologie und Intensivmedizin, Universität Duisburg-Essen, Universitätsklinikum Essen, 45122 Essen, GermanyKlinik für Anästhesiologie und Intensivmedizin, Universität Duisburg-Essen, Universitätsklinikum Essen, 45122 Essen, GermanyInstitut für Biochemie/CCM, Charité-Universitätsmedizin-Berlin, 13347 Berlin, GermanyKlinik für Pneumologie und Allergologie, Ruhrlandklinik, Universität Duisburg-Essen, 45239 Essen, GermanyMax Planck Institute for Infection Biology, Core Facility Protein Analysis, 13125 Berlin, GermanyInstitut für Biochemie/CCM, Charité-Universitätsmedizin-Berlin, 13347 Berlin, GermanyKlinik für Anästhesiologie und Intensivmedizin, Universität Duisburg-Essen, Universitätsklinikum Essen, 45122 Essen, GermanyThere is increasing evidence that proteasomes have a biological role in the extracellular alveolar space, but inflammation could change their composition. We tested whether immunoproteasome protein-containing subpopulations are present in the alveolar space of patients with lung inflammation evoking the acute respiratory distress syndrome (ARDS). Bronchoalveolar lavage (BAL) supernatants and cell pellet lysate from ARDS patients (n=28) and healthy subjects (n=10) were analyzed for the presence of immunoproteasome proteins (LMP2 and LMP7) and proteasome subtypes by western blot, chromatographic purification, and 2D-dimensional gelelectrophoresis. In all ARDS patients but not in healthy subjects LMP7 and LMP2 were observed in BAL supernatants. Proteasomes purified from pooled ARDS BAL supernatant showed an altered enzyme activity ratio. Chromatography revealed a distinct pattern with 7 proteasome subtype peaks in BAL supernatant of ARDS patients that differed from healthy subjects. Total proteasome concentration in BAL supernatant was increased in ARDS (971 ng/mL ± 1116 versus 59±25; P<0.001), and all fluorogenic substrates were hydrolyzed, albeit to a lesser extent, with inhibition by epoxomicin (P=0.0001). Thus, we identified for the first time immunoproteasome proteins and a distinct proteasomal subtype pattern in the alveolar space of ARDS patients, presumably in response to inflammation.http://dx.doi.org/10.1155/2012/204250 |
| spellingShingle | S. U. Sixt R. Alami J. Hakenbeck M. Adamzik A. Kloß U. Costabel P. R. Jungblut B. Dahlmann J. Peters Distinct Proteasome Subpopulations in the Alveolar Space of Patients with the Acute Respiratory Distress Syndrome Mediators of Inflammation |
| title | Distinct Proteasome Subpopulations in the Alveolar Space of Patients with the Acute Respiratory Distress Syndrome |
| title_full | Distinct Proteasome Subpopulations in the Alveolar Space of Patients with the Acute Respiratory Distress Syndrome |
| title_fullStr | Distinct Proteasome Subpopulations in the Alveolar Space of Patients with the Acute Respiratory Distress Syndrome |
| title_full_unstemmed | Distinct Proteasome Subpopulations in the Alveolar Space of Patients with the Acute Respiratory Distress Syndrome |
| title_short | Distinct Proteasome Subpopulations in the Alveolar Space of Patients with the Acute Respiratory Distress Syndrome |
| title_sort | distinct proteasome subpopulations in the alveolar space of patients with the acute respiratory distress syndrome |
| url | http://dx.doi.org/10.1155/2012/204250 |
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