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Shotgun proteomic analysis of human induced sputum

Shotgun proteomic analysis of human induced sputum
Shotgun proteomic analysis of human induced sputum
Sputum is increasingly recognised as a biologically relevant sample of the environment of the airways when collected under controlled conditions. As such, it has been used for the analysis of bacterial load, tumour and inflammatory cell content, and of biomarkers of airways diseases such as cystic fibrosis, asthma and chronic obstructive pulmonary disease. While related biofluids such as broncho-alveolar lavage1 fluid, nasal lavage fluid,2 and in particular saliva3 have been well-characterised using standard 2-D gel based proteomics, sputum has not. This is probably due to the presence of mucin glycoproteins that compromise the separation of proteins during the IEF stage of 2-D gel analysis, preventing good resolution.
We have used a combination of 2-D gel electrophoresis (fig 1BGo) and a multidimensional mass-spectrometric technique (GeLC-MS/MS) (fig 1AGo) to examine the proteome of induced sputum. 191 individual human proteins were confidently assigned using these techniques. In addition to expected components, several hitherto unreported proteins were discovered, including three members of the annexin family, kallikreins 1 and 11, and peroxiredoxins 1, 2, and 5. The resultant data represent the first detailed survey of the human induced sputum proteome and allow a comparison of the induced sputum proteome with other biologically related fluids such as broncho-alveolar lavage fluid and saliva. They also provide a platform for the future identification of biomarkers of airways disease.
0040-6376
P206
Nicholas, B.
785c44fb-6536-4189-803b-4545425e9385
Skipp, P.
1ba7dcf6-9fe7-4b5c-a9d0-e32ed7f42aa5
Mould, R.
cf0b5fe6-be7f-4dbe-a709-5c6271f1624f
Rennard, S.
5ed888ed-6c3d-47d8-8535-99089c150c0b
Davies, D.E.
7de8fdc7-3640-4e3a-aa91-d0e03f990c38
Connor, D.
0f9b0290-24f9-406f-961e-724f5de4a0fe
Djukanovic, R.
d9a45ee7-6a80-4d84-a0ed-10962660a98d
Nicholas, B.
785c44fb-6536-4189-803b-4545425e9385
Skipp, P.
1ba7dcf6-9fe7-4b5c-a9d0-e32ed7f42aa5
Mould, R.
cf0b5fe6-be7f-4dbe-a709-5c6271f1624f
Rennard, S.
5ed888ed-6c3d-47d8-8535-99089c150c0b
Davies, D.E.
7de8fdc7-3640-4e3a-aa91-d0e03f990c38
Connor, D.
0f9b0290-24f9-406f-961e-724f5de4a0fe
Djukanovic, R.
d9a45ee7-6a80-4d84-a0ed-10962660a98d

Nicholas, B., Skipp, P., Mould, R., Rennard, S., Davies, D.E., Connor, D. and Djukanovic, R. (2006) Shotgun proteomic analysis of human induced sputum. Thorax, 61 (Supp/2), P206.

Record type: Article

Abstract

Sputum is increasingly recognised as a biologically relevant sample of the environment of the airways when collected under controlled conditions. As such, it has been used for the analysis of bacterial load, tumour and inflammatory cell content, and of biomarkers of airways diseases such as cystic fibrosis, asthma and chronic obstructive pulmonary disease. While related biofluids such as broncho-alveolar lavage1 fluid, nasal lavage fluid,2 and in particular saliva3 have been well-characterised using standard 2-D gel based proteomics, sputum has not. This is probably due to the presence of mucin glycoproteins that compromise the separation of proteins during the IEF stage of 2-D gel analysis, preventing good resolution.
We have used a combination of 2-D gel electrophoresis (fig 1BGo) and a multidimensional mass-spectrometric technique (GeLC-MS/MS) (fig 1AGo) to examine the proteome of induced sputum. 191 individual human proteins were confidently assigned using these techniques. In addition to expected components, several hitherto unreported proteins were discovered, including three members of the annexin family, kallikreins 1 and 11, and peroxiredoxins 1, 2, and 5. The resultant data represent the first detailed survey of the human induced sputum proteome and allow a comparison of the induced sputum proteome with other biologically related fluids such as broncho-alveolar lavage fluid and saliva. They also provide a platform for the future identification of biomarkers of airways disease.

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Published date: 1 December 2006

Identifiers

Local EPrints ID: 56006
URI: http://eprints.soton.ac.uk/id/eprint/56006
ISSN: 0040-6376
PURE UUID: 05d50b5b-a595-43de-b271-71f34834b39a
ORCID for B. Nicholas: ORCID iD orcid.org/0000-0003-1467-9643
ORCID for P. Skipp: ORCID iD orcid.org/0000-0002-2995-2959
ORCID for D.E. Davies: ORCID iD orcid.org/0000-0002-5117-2991
ORCID for R. Djukanovic: ORCID iD orcid.org/0000-0001-6039-5612

Catalogue record

Date deposited: 07 Aug 2008
Last modified: 18 Mar 2024 02:58

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Contributors

Author: B. Nicholas ORCID iD
Author: P. Skipp ORCID iD
Author: R. Mould
Author: S. Rennard
Author: D.E. Davies ORCID iD
Author: D. Connor
Author: R. Djukanovic ORCID iD

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