Identification of Fibrin Clot-Bound Plasma Proteins
Open Access
- 3 August 2012
- journal article
- research article
- Published by Public Library of Science (PLoS) in PLOS ONE
- Vol. 7 (8), e41966
- https://doi.org/10.1371/journal.pone.0041966
Abstract
Several proteins are known to bind to a fibrin network and to change clot properties or function. In this study we aimed to get an overview of fibrin clot-bound plasma proteins. A plasma clot was formed by adding thrombin, CaCl2 and aprotinin to citrated platelet-poor plasma and unbound proteins were washed away with Tris-buffered saline. Non-covalently bound proteins were extracted, separated with 2D gel electrophoresis and visualized with Sypro Ruby. Excised protein spots were analyzed with mass spectrometry. The identity of the proteins was verified by checking the mass of the protein, and, if necessary, by Western blot analysis. Next to established fibrin-binding proteins we identified several novel fibrin clot-bound plasma proteins, including α2-macroglobulin, carboxypeptidase N, α1-antitrypsin, haptoglobin, serum amyloid P, and the apolipoproteins A-I, E, J, and A-IV. The latter six proteins are associated with high-density lipoprotein particles. In addition we showed that high-density lipoprotein associated proteins were also present in fibrinogen preparations purified from plasma. Most plasma proteins in a fibrin clot can be classified into three groups according to either blood coagulation, protease inhibition or high-density lipoprotein metabolism. The presence of high-density lipoprotein in clots might point to a role in hemostasis.Keywords
This publication has 46 references indexed in Scilit:
- Reevaluation of the role of HDL in the anticoagulant activated protein C system in humansJCI Insight, 2010
- Haptoglobin inhibits phospholipid transfer protein activity in hyperlipidemic human plasmaLipids in Health and Disease, 2009
- Anionic Phospholipids Lose Their Procoagulant Properties When Incorporated into High Density LipoproteinsJournal of Biological Chemistry, 2009
- Structure of fibrin: impact on clot stabilityJournal of Thrombosis and Haemostasis, 2007
- Shotgun proteomics implicates protease inhibition and complement activation in the antiinflammatory properties of HDLJCI Insight, 2007
- Hepatocyte-derived fibrinogen-related protein-1 is associated with the fibrin matrix of a plasma clotBiochemical and Biophysical Research Communications, 2006
- Current two‐dimensional electrophoresis technology for proteomicsProteomics, 2004
- α2‐Macroglobulin: A Sensor for ProteolysisAnnals of the New York Academy of Sciences, 1994
- Identification of Proteins Associated with Apolipoprotein A-I-Containing Lipoproteins Purified by Selected-Affinity ImmunosorptionBiochemistry, 1994
- Molecular basis of fibrinogen Naples associated with defective thrombin binding and thrombophilia. Homozygous substitution of B beta 68 Ala----Thr.JCI Insight, 1992