Advanced

Gla-domain mutated human protein C exhibiting enhanced anticoagulant activity and increased phospholipid binding.

Sun, Yong-Hui; Shen, Lei and Dahlbäck, Björn LU (2003) In Blood 101(6). p.2277-2284
Abstract
Protein C is a member of the vitamin K-dependent protein family. Proteins in this family have similar gamma-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 (OGN), 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 (WTAPC), while QGN-APC and SEDY-APC did not. Both normal... (More)
Protein C is a member of the vitamin K-dependent protein family. Proteins in this family have similar gamma-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 (OGN), 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 (WTAPC), 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 phospolipid membranes. The unique properties of QGNSEDY-APC suggest this APC variant possibly to have greater therapeutic potential than WT APC. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Blood
volume
101
issue
6
pages
2277 - 2284
publisher
American Society of Hematology
external identifiers
  • pmid:12446455
  • wos:000181432600032
  • scopus:0037443444
ISSN
1528-0020
DOI
10.1182/blood-2002-06-1691
language
English
LU publication?
yes
id
bb2622a9-0e1c-4e46-bb81-d6977d60aa2f (old id 111014)
alternative location
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12446455&dopt=Abstract
date added to LUP
2007-07-27 10:52:03
date last changed
2018-10-03 10:15:40
@article{bb2622a9-0e1c-4e46-bb81-d6977d60aa2f,
  abstract     = {Protein C is a member of the vitamin K-dependent protein family. Proteins in this family have similar gamma-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 (OGN), 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 (WTAPC), 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 phospolipid membranes. The unique properties of QGNSEDY-APC suggest this APC variant possibly to have greater therapeutic potential than WT APC.},
  author       = {Sun, Yong-Hui and Shen, Lei and Dahlbäck, Björn},
  issn         = {1528-0020},
  language     = {eng},
  number       = {6},
  pages        = {2277--2284},
  publisher    = {American Society of Hematology},
  series       = {Blood},
  title        = {Gla-domain mutated human protein C exhibiting enhanced anticoagulant activity and increased phospholipid binding.},
  url          = {http://dx.doi.org/10.1182/blood-2002-06-1691},
  volume       = {101},
  year         = {2003},
}