High throughput proteomic analysis of the secretome in an explant model of articular cartilage inflammation.

Clutterbuck, AL, Smith, JR, Allaway, D, Harris, P, Liddell, S and Mobasheri, A (2011) High throughput proteomic analysis of the secretome in an explant model of articular cartilage inflammation. J Proteomics, 74 (5). pp. 704-715.

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Abstract

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, 10ng/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.

Item Type:	Article
Authors :	Name Email ORCID Clutterbuck, AL Smith, JR Allaway, D Harris, P Liddell, S Mobasheri, A a.mobasheri@surrey.ac.uk
Date :	1 May 2011
Identification Number :	10.1016/j.jprot.2011.02.017
Uncontrolled Keywords :	Animals, Anti-Inflammatory Agents, Non-Steroidal, Carbazoles, Cartilage, Articular, Gene Expression Regulation, Horses, Inflammation, Models, Biological, Osteochondritis, Proteome
Related URLs :	Author
Depositing User :	Symplectic Elements
Date Deposited :	17 May 2017 10:11
Last Modified :	17 May 2017 14:47
URI:	http://epubs.surrey.ac.uk/id/eprint/826776

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