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Proteomics Strategy for Identifying Candidate Bioactive Proteins in Complex Mixtures : Application to the Platelet Releasate
Joint Authors
Cagney, Gerard
O'Connor, Roisin
O'Brien, Colm
de Stefani, Andreas
Fitzgerald, Desmond
Cryan, Lorna M.
Wynne, Kieran
Source
Journal of Biomedicine and Biotechnology
Issue
Vol. 2010, Issue 2010 (31 Dec. 2010), pp.1-12, 12 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2010-03-24
Country of Publication
Egypt
No. of Pages
12
Main Subjects
Abstract EN
Proteomic approaches have proven powerful at identifying large numbers of proteins, but there are fewer reports of functional characterization of proteins in biological tissues.
Here, we describe an experimental approach that fractionates proteins released from human platelets, linking bioassay activity to identity.
We used consecutive orthogonal separation platforms to ensure sensitive detection: (a) ion-exchange of intact proteins, (b) SDS-PAGE separation of ion-exchange fractions and (c) HPLC separation of tryptic digests coupled to electrospray tandem mass spectrometry.
Migration of THP-1 monocytes in response to complete or fractionated platelet releasate was assessed and located to just one of the forty-nine ion-exchange fractions.
Over 300 proteins were identified in the releasate, with a wide range of annotated biophysical and biochemical properties, in particular platelet activation, adhesion, and wound healing.
The presence of PEDF and involucrin, two proteins not previously reported in platelet releasate, was confirmed by western blotting.
Proteins identified within the fraction with monocyte promigratory activity and not in other inactive fractions included vimentin, PEDF, and TIMP-1.
We conclude that this analytical platform is effective for the characterization of complex bioactive samples.
American Psychological Association (APA)
O'Connor, Roisin& Cryan, Lorna M.& Wynne, Kieran& de Stefani, Andreas& Fitzgerald, Desmond& O'Brien, Colm…[et al.]. 2010. Proteomics Strategy for Identifying Candidate Bioactive Proteins in Complex Mixtures : Application to the Platelet Releasate. Journal of Biomedicine and Biotechnology،Vol. 2010, no. 2010, pp.1-12.
https://search.emarefa.net/detail/BIM-447002
Modern Language Association (MLA)
O'Connor, Roisin…[et al.]. Proteomics Strategy for Identifying Candidate Bioactive Proteins in Complex Mixtures : Application to the Platelet Releasate. Journal of Biomedicine and Biotechnology No. 2010 (2010), pp.1-12.
https://search.emarefa.net/detail/BIM-447002
American Medical Association (AMA)
O'Connor, Roisin& Cryan, Lorna M.& Wynne, Kieran& de Stefani, Andreas& Fitzgerald, Desmond& O'Brien, Colm…[et al.]. Proteomics Strategy for Identifying Candidate Bioactive Proteins in Complex Mixtures : Application to the Platelet Releasate. Journal of Biomedicine and Biotechnology. 2010. Vol. 2010, no. 2010, pp.1-12.
https://search.emarefa.net/detail/BIM-447002
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-447002