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Gla domain–mutated human protein C exhibiting enhanced anticoagulant activity and increased phospholipid binding
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| Zeitschriftentitel: | Blood |
|---|---|
| Personen und Körperschaften: | , , |
| In: | Blood, 101, 2003, 6, S. 2277-2284 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
American Society of Hematology
|
| Schlagwörter: |
| author_facet |
Sun, Yong-Hui Shen, Lei Dahlbäck, Björn Sun, Yong-Hui Shen, Lei Dahlbäck, Björn |
|---|---|
| author |
Sun, Yong-Hui Shen, Lei Dahlbäck, Björn |
| spellingShingle |
Sun, Yong-Hui Shen, Lei Dahlbäck, Björn Blood Gla domain–mutated human protein C exhibiting enhanced anticoagulant activity and increased phospholipid binding Cell Biology Hematology Immunology Biochemistry |
| author_sort |
sun, yong-hui |
| spelling |
Sun, Yong-Hui Shen, Lei Dahlbäck, Björn 1528-0020 0006-4971 American Society of Hematology Cell Biology Hematology Immunology Biochemistry http://dx.doi.org/10.1182/blood-2002-06-1691 <jats:p>Protein C is a member of the vitamin K– dependent protein family. Proteins in this family have similar γ-carboxyglutamic acid (Gla)–rich domains, but their affinities for negatively charged phospholipid membranes vary more than 1000-fold. We have shown that it is possible to enhance anticoagulant activity and membrane affinity of protein C by selective mutagenesis of the Gla domain. In this study, 3 new mutants, Q10G11N12 (QGN), S23E32D33Y44 (SEDY), and Q10G11N12S23E32D33Y44 (QGNSEDY), were created. In plasma-based coagulation assays, the activated form of QGNSEDY (QGNSEDY-APC) demonstrated approximately 20-fold higher anticoagulant activity than wild-type activated protein C (WT APC), while QGN-APC and SEDY-APC did not. Both normal activated factor V (FVa) and FVa Leiden (Arg506Gln) were degraded much more efficiently by QGNSEDY-APC than by WT APC in the presence as well as in the absence of protein S. Binding of protein C variants to negatively charged phospholipid membranes was investigated using light scattering and the BIAcore technique. QGNSEDY demonstrated 3- to 7-fold enhanced binding as compared with WT protein C, suggesting the membrane affinity to be influenced by several residues located at different parts of the Gla domain. The anticoagulant activity as well as phospholipid binding ability was only enhanced when multiple regions of the Gla domain were modified. The results provide insights into the molecular mechanisms that are involved in determining the binding affinity of the interaction between Gla domains and phospholipid membranes. The unique properties of QGNSEDY-APC suggest this APC variant possibly to have greater therapeutic potential than WT APC.</jats:p> Gla domain–mutated human protein C exhibiting enhanced anticoagulant activity and increased phospholipid binding Blood |
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10.1182/blood-2002-06-1691 |
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DE-Gla1 DE-Zi4 DE-15 DE-Pl11 DE-Rs1 DE-105 DE-14 DE-Ch1 DE-L229 DE-D275 DE-Bn3 DE-Brt1 DE-Zwi2 DE-D161 |
| imprint |
American Society of Hematology, 2003 |
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American Society of Hematology, 2003 |
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1528-0020 0006-4971 |
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1528-0020 0006-4971 |
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2003 |
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American Society of Hematology |
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49 |
| title |
Gla domain–mutated human protein C exhibiting enhanced anticoagulant activity and increased phospholipid binding |
| title_unstemmed |
Gla domain–mutated human protein C exhibiting enhanced anticoagulant activity and increased phospholipid binding |
| title_full |
Gla domain–mutated human protein C exhibiting enhanced anticoagulant activity and increased phospholipid binding |
| title_fullStr |
Gla domain–mutated human protein C exhibiting enhanced anticoagulant activity and increased phospholipid binding |
| title_full_unstemmed |
Gla domain–mutated human protein C exhibiting enhanced anticoagulant activity and increased phospholipid binding |
| title_short |
Gla domain–mutated human protein C exhibiting enhanced anticoagulant activity and increased phospholipid binding |
| title_sort |
gla domain–mutated human protein c exhibiting enhanced anticoagulant activity and increased phospholipid binding |
| topic |
Cell Biology Hematology Immunology Biochemistry |
| url |
http://dx.doi.org/10.1182/blood-2002-06-1691 |
| publishDate |
2003 |
| physical |
2277-2284 |
| description |
<jats:p>Protein C is a member of the vitamin K– dependent protein family. Proteins in this family have similar γ-carboxyglutamic acid (Gla)–rich domains, but their affinities for negatively charged phospholipid membranes vary more than 1000-fold. We have shown that it is possible to enhance anticoagulant activity and membrane affinity of protein C by selective mutagenesis of the Gla domain. In this study, 3 new mutants, Q10G11N12 (QGN), S23E32D33Y44 (SEDY), and Q10G11N12S23E32D33Y44 (QGNSEDY), were created. In plasma-based coagulation assays, the activated form of QGNSEDY (QGNSEDY-APC) demonstrated approximately 20-fold higher anticoagulant activity than wild-type activated protein C (WT APC), while QGN-APC and SEDY-APC did not. Both normal activated factor V (FVa) and FVa Leiden (Arg506Gln) were degraded much more efficiently by QGNSEDY-APC than by WT APC in the presence as well as in the absence of protein S. Binding of protein C variants to negatively charged phospholipid membranes was investigated using light scattering and the BIAcore technique. QGNSEDY demonstrated 3- to 7-fold enhanced binding as compared with WT protein C, suggesting the membrane affinity to be influenced by several residues located at different parts of the Gla domain. The anticoagulant activity as well as phospholipid binding ability was only enhanced when multiple regions of the Gla domain were modified. The results provide insights into the molecular mechanisms that are involved in determining the binding affinity of the interaction between Gla domains and phospholipid membranes. The unique properties of QGNSEDY-APC suggest this APC variant possibly to have greater therapeutic potential than WT APC.</jats:p> |
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| author | Sun, Yong-Hui, Shen, Lei, Dahlbäck, Björn |
| author_facet | Sun, Yong-Hui, Shen, Lei, Dahlbäck, Björn, Sun, Yong-Hui, Shen, Lei, Dahlbäck, Björn |
| author_sort | sun, yong-hui |
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| description | <jats:p>Protein C is a member of the vitamin K– dependent protein family. Proteins in this family have similar γ-carboxyglutamic acid (Gla)–rich domains, but their affinities for negatively charged phospholipid membranes vary more than 1000-fold. We have shown that it is possible to enhance anticoagulant activity and membrane affinity of protein C by selective mutagenesis of the Gla domain. In this study, 3 new mutants, Q10G11N12 (QGN), S23E32D33Y44 (SEDY), and Q10G11N12S23E32D33Y44 (QGNSEDY), were created. In plasma-based coagulation assays, the activated form of QGNSEDY (QGNSEDY-APC) demonstrated approximately 20-fold higher anticoagulant activity than wild-type activated protein C (WT APC), while QGN-APC and SEDY-APC did not. Both normal activated factor V (FVa) and FVa Leiden (Arg506Gln) were degraded much more efficiently by QGNSEDY-APC than by WT APC in the presence as well as in the absence of protein S. Binding of protein C variants to negatively charged phospholipid membranes was investigated using light scattering and the BIAcore technique. QGNSEDY demonstrated 3- to 7-fold enhanced binding as compared with WT protein C, suggesting the membrane affinity to be influenced by several residues located at different parts of the Gla domain. The anticoagulant activity as well as phospholipid binding ability was only enhanced when multiple regions of the Gla domain were modified. The results provide insights into the molecular mechanisms that are involved in determining the binding affinity of the interaction between Gla domains and phospholipid membranes. The unique properties of QGNSEDY-APC suggest this APC variant possibly to have greater therapeutic potential than WT APC.</jats:p> |
| doi_str_mv | 10.1182/blood-2002-06-1691 |
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| imprint | American Society of Hematology, 2003 |
| imprint_str_mv | American Society of Hematology, 2003 |
| institution | DE-Gla1, DE-Zi4, DE-15, DE-Pl11, DE-Rs1, DE-105, DE-14, DE-Ch1, DE-L229, DE-D275, DE-Bn3, DE-Brt1, DE-Zwi2, DE-D161 |
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| language | English |
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| spelling | Sun, Yong-Hui Shen, Lei Dahlbäck, Björn 1528-0020 0006-4971 American Society of Hematology Cell Biology Hematology Immunology Biochemistry http://dx.doi.org/10.1182/blood-2002-06-1691 <jats:p>Protein C is a member of the vitamin K– dependent protein family. Proteins in this family have similar γ-carboxyglutamic acid (Gla)–rich domains, but their affinities for negatively charged phospholipid membranes vary more than 1000-fold. We have shown that it is possible to enhance anticoagulant activity and membrane affinity of protein C by selective mutagenesis of the Gla domain. In this study, 3 new mutants, Q10G11N12 (QGN), S23E32D33Y44 (SEDY), and Q10G11N12S23E32D33Y44 (QGNSEDY), were created. In plasma-based coagulation assays, the activated form of QGNSEDY (QGNSEDY-APC) demonstrated approximately 20-fold higher anticoagulant activity than wild-type activated protein C (WT APC), while QGN-APC and SEDY-APC did not. Both normal activated factor V (FVa) and FVa Leiden (Arg506Gln) were degraded much more efficiently by QGNSEDY-APC than by WT APC in the presence as well as in the absence of protein S. Binding of protein C variants to negatively charged phospholipid membranes was investigated using light scattering and the BIAcore technique. QGNSEDY demonstrated 3- to 7-fold enhanced binding as compared with WT protein C, suggesting the membrane affinity to be influenced by several residues located at different parts of the Gla domain. The anticoagulant activity as well as phospholipid binding ability was only enhanced when multiple regions of the Gla domain were modified. The results provide insights into the molecular mechanisms that are involved in determining the binding affinity of the interaction between Gla domains and phospholipid membranes. The unique properties of QGNSEDY-APC suggest this APC variant possibly to have greater therapeutic potential than WT APC.</jats:p> Gla domain–mutated human protein C exhibiting enhanced anticoagulant activity and increased phospholipid binding Blood |
| spellingShingle | Sun, Yong-Hui, Shen, Lei, Dahlbäck, Björn, Blood, Gla domain–mutated human protein C exhibiting enhanced anticoagulant activity and increased phospholipid binding, Cell Biology, Hematology, Immunology, Biochemistry |
| title | Gla domain–mutated human protein C exhibiting enhanced anticoagulant activity and increased phospholipid binding |
| title_full | Gla domain–mutated human protein C exhibiting enhanced anticoagulant activity and increased phospholipid binding |
| title_fullStr | Gla domain–mutated human protein C exhibiting enhanced anticoagulant activity and increased phospholipid binding |
| title_full_unstemmed | Gla domain–mutated human protein C exhibiting enhanced anticoagulant activity and increased phospholipid binding |
| title_short | Gla domain–mutated human protein C exhibiting enhanced anticoagulant activity and increased phospholipid binding |
| title_sort | gla domain–mutated human protein c exhibiting enhanced anticoagulant activity and increased phospholipid binding |
| title_unstemmed | Gla domain–mutated human protein C exhibiting enhanced anticoagulant activity and increased phospholipid binding |
| topic | Cell Biology, Hematology, Immunology, Biochemistry |
| url | http://dx.doi.org/10.1182/blood-2002-06-1691 |