| Title: |
Protein S multimers are generated in vitro and affect protein S structure-function analyses |
| Authors: |
Sere, KM; Willems, GM; Rosing, J; Hackeng, TM |
| Source: |
Sere, KM, Willems, GM, Rosing, J & Hackeng, TM 2006, 'Protein S multimers are generated in vitro and affect protein S structure-function analyses', Paper presented at 20th Congress of the International-Society-on-Thrombosis-and-Haemostasis, Sydney, Australia, 6/08/05 - 12/08/05 pp. S111-S120. https://doi.org/10.1053/j.seminhematol.2005.11.027 |
| Publication Year: |
2006 |
| Collection: |
Maastricht University Research Publications |
| Subject Terms: |
HIGH-AFFINITY INTERACTION; HUMAN FACTOR-VIII; C4B-BINDING PROTEIN; FACTOR-VA; COFACTOR ACTIVITY; PROTHROMBINASE COMPLEX; ENDOTHELIAL-CELLS; FACTOR-XA; BINDING; INHIBITION |
| Description: |
Purified human protein S preparations contain small amounts of multimeric protein S. Protein S multimers are absent in plasma, suggesting that multimerization results from purification. Protein S multimers effectively inhibit phospholipid-dependent reactions at low phospholipid concentrations, and may therefore interfere during functional analysis of protein S. We have demonstrated that anion-exchange chromatography, as well as high ionic strength or low pH elution conditions used in immunoaffinity purification of protein S, induce protein S multimer formation. When protein S multimers were removed from protein S preparations by size-exclusion chromatography, multimers spontaneously reappeared in the protein S monomer fraction. In model systems, high phospholipid concentrations (>50 micromol/L) completely abrogate the inhibitory effect of protein S multimers on prothrombinase complex activity. In addition, C4BP does not bind to protein S multimers. Thus, at low phospholipid concentrations, addition of C4BP to purified protein S will not affect the inhibitory activity of protein S multimers. In conclusion, to avoid misinterpretations during protein S structure-function analysis due to multimers present in purified protein S preparations, we recommend studying the anticoagulant activities of protein S either in plasma, where protein S is in its unmodified natural form, or at high phospholipid concentrations in model systems with purified proteins. |
| Document Type: |
conference object |
| Language: |
English |
| Relation: |
info:eu-repo/semantics/altIdentifier/wos/000234980100021 |
| DOI: |
10.1053/j.seminhematol.2005.11.027 |
| Availability: |
https://cris.maastrichtuniversity.nl/en/publications/f7c9b616-9a60-4f18-955a-43f71dcdf4c5; https://doi.org/10.1053/j.seminhematol.2005.11.027 |
| Rights: |
info:eu-repo/semantics/closedAccess |
| Accession Number: |
edsbas.5FF69332 |
| Database: |
BASE |