High throughput proteomic analysis of the secretome in an explant model of articular cartilage inflammation
This study employed a targeted high-throughput proteomic approach to identify the major proteins present in the secretome of articular cartilage. Explants from equine metacarpophalangeal joints were incubated alone or with interleukin-1beta (IL-1β, 10 ng/ml), with or without carprofen, a non-steroid...
| Main Authors: | , , , , , |
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| Format: | Article |
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Elsevier
2011
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| Online Access: | https://eprints.nottingham.ac.uk/2298/ |
| _version_ | 1848790749510893568 |
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| author | Clutterbuck, Abigail L. Smith, Julia R. Allaway, David Harris, Pat Liddell, Susan Mobasheri, Ali |
| author_facet | Clutterbuck, Abigail L. Smith, Julia R. Allaway, David Harris, Pat Liddell, Susan Mobasheri, Ali |
| author_sort | Clutterbuck, Abigail L. |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | This study employed a targeted high-throughput proteomic approach to identify the major proteins present in the secretome of articular cartilage. Explants from equine metacarpophalangeal joints were incubated alone or with interleukin-1beta (IL-1β, 10 ng/ml), with or without carprofen, a non-steroidal anti-inflammatory drug, for six days. After tryptic digestion of culture medium supernatants, resulting peptides were separated by HPLC and detected in a Bruker amaZon ion trap instrument. The five most abundant peptides in each MS scan were fragmented and the fragmentation patterns compared to mammalian entries in the Swiss-Prot database, using the Mascot search engine. Tryptic peptides originating from aggrecan core protein, cartilage oligomeric matrix protein (COMP), fibronectin, fibromodulin, thrombospondin-1 (TSP-1), clusterin (CLU), cartilage intermediate layer protein-1 (CILP-1), chondroadherin (CHAD) and matrix metalloproteinases MMP-1 and MMP-3 were detected. Quantitative western blotting confirmed the presence of CILP-1, CLU, MMP-1, MMP-3 and TSP-1. Treatment with IL-1β increased MMP-1, MMP-3 and TSP-1 and decreased the CLU precursor but did not affect CILP-1 and CLU levels. Many of the proteins identified have well-established extracellular matrix functions and are involved in early repair/stress responses in cartilage. This high throughput approach may be used to study the changes that occur in the early stages of osteoarthritis. |
| first_indexed | 2025-11-14T18:17:34Z |
| format | Article |
| id | nottingham-2298 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T18:17:34Z |
| publishDate | 2011 |
| publisher | Elsevier |
| recordtype | eprints |
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| spelling | nottingham-22982020-05-04T16:30:31Z https://eprints.nottingham.ac.uk/2298/ High throughput proteomic analysis of the secretome in an explant model of articular cartilage inflammation Clutterbuck, Abigail L. Smith, Julia R. Allaway, David Harris, Pat Liddell, Susan Mobasheri, Ali This study employed a targeted high-throughput proteomic approach to identify the major proteins present in the secretome of articular cartilage. Explants from equine metacarpophalangeal joints were incubated alone or with interleukin-1beta (IL-1β, 10 ng/ml), with or without carprofen, a non-steroidal anti-inflammatory drug, for six days. After tryptic digestion of culture medium supernatants, resulting peptides were separated by HPLC and detected in a Bruker amaZon ion trap instrument. The five most abundant peptides in each MS scan were fragmented and the fragmentation patterns compared to mammalian entries in the Swiss-Prot database, using the Mascot search engine. Tryptic peptides originating from aggrecan core protein, cartilage oligomeric matrix protein (COMP), fibronectin, fibromodulin, thrombospondin-1 (TSP-1), clusterin (CLU), cartilage intermediate layer protein-1 (CILP-1), chondroadherin (CHAD) and matrix metalloproteinases MMP-1 and MMP-3 were detected. Quantitative western blotting confirmed the presence of CILP-1, CLU, MMP-1, MMP-3 and TSP-1. Treatment with IL-1β increased MMP-1, MMP-3 and TSP-1 and decreased the CLU precursor but did not affect CILP-1 and CLU levels. Many of the proteins identified have well-established extracellular matrix functions and are involved in early repair/stress responses in cartilage. This high throughput approach may be used to study the changes that occur in the early stages of osteoarthritis. Elsevier 2011-05-01 Article PeerReviewed Clutterbuck, Abigail L., Smith, Julia R., Allaway, David, Harris, Pat, Liddell, Susan and Mobasheri, Ali (2011) High throughput proteomic analysis of the secretome in an explant model of articular cartilage inflammation. Journal of Proteomics, 74 (5). pp. 704-715. ISSN 1874-3919 http://www.sciencedirect.com/science/article/pii/S1874391911000558 doi:10.1016/j.jprot.2011.02.017 doi:10.1016/j.jprot.2011.02.017 |
| spellingShingle | Clutterbuck, Abigail L. Smith, Julia R. Allaway, David Harris, Pat Liddell, Susan Mobasheri, Ali High throughput proteomic analysis of the secretome in an explant model of articular cartilage inflammation |
| title | High throughput proteomic analysis of the secretome in an explant model of articular cartilage inflammation |
| title_full | High throughput proteomic analysis of the secretome in an explant model of articular cartilage inflammation |
| title_fullStr | High throughput proteomic analysis of the secretome in an explant model of articular cartilage inflammation |
| title_full_unstemmed | High throughput proteomic analysis of the secretome in an explant model of articular cartilage inflammation |
| title_short | High throughput proteomic analysis of the secretome in an explant model of articular cartilage inflammation |
| title_sort | high throughput proteomic analysis of the secretome in an explant model of articular cartilage inflammation |
| url | https://eprints.nottingham.ac.uk/2298/ https://eprints.nottingham.ac.uk/2298/ https://eprints.nottingham.ac.uk/2298/ |