Activated protein C cofactor function of protein S: a critical role for Asp95 in the EGF1-like domain
(2010) In Blood 115(23). p.4878-4885- Abstract
- Protein S has an established role in the protein C anticoagulant pathway, where it enhances the factor Va (FVa) and factor VIIIa (FVIIIa) inactivating property of activated protein C (APC). Despite its physiological role and clinical importance, the molecular basis of its action is not fully understood. To clarify the mechanism of the protein S interaction with APC, we have constructed and expressed a library of composite or point variants of human protein S, with residue substitutions introduced into the Gla, thrombin-sensitive region (TSR), epidermal growth factor 1 (EGF1), and EGF2 domains. Cofactor activity for APC was evaluated by calibrated automated thrombography (CAT) using protein S-deficient plasma. Of 27 variants tested... (More)
- Protein S has an established role in the protein C anticoagulant pathway, where it enhances the factor Va (FVa) and factor VIIIa (FVIIIa) inactivating property of activated protein C (APC). Despite its physiological role and clinical importance, the molecular basis of its action is not fully understood. To clarify the mechanism of the protein S interaction with APC, we have constructed and expressed a library of composite or point variants of human protein S, with residue substitutions introduced into the Gla, thrombin-sensitive region (TSR), epidermal growth factor 1 (EGF1), and EGF2 domains. Cofactor activity for APC was evaluated by calibrated automated thrombography (CAT) using protein S-deficient plasma. Of 27 variants tested initially, only one, protein S D95A (within the EGF1 domain), was largely devoid of functional APC cofactor activity. Protein S D95A was, however, gamma-carboxylated and bound phospholipids with an apparent dissociation constant (Kd(app)) similar to that of wildtype (WT) protein S. In a purified assay using FVa R506Q/R679Q, purified protein S D95A was shown to have greatly reduced ability to enhance APC-induced cleavage of FVa Arg306. It is concluded that residue Asp95 within EGF1 is critical forAPC cofactor function of protein S and could define a principal functional interaction site for APC. (Blood. 2010;115(23):4878-4885) (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1760680
- author
- Andersson, Helena M. ; Arantes, Marcia J. ; Crawley, James T. B. ; Luken, Brenda M. ; Tran, Sinh LU ; Dahlbäck, Björn LU ; Lane, David A. and Rezende, Suely M.
- organization
- publishing date
- 2010
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Blood
- volume
- 115
- issue
- 23
- pages
- 4878 - 4885
- publisher
- American Society of Hematology
- external identifiers
-
- wos:000278635900036
- scopus:77954707351
- pmid:20308596
- ISSN
- 1528-0020
- DOI
- 10.1182/blood-2009-11-256610
- language
- English
- LU publication?
- yes
- id
- 1fdd62b1-df73-485c-b248-01dd724c47b7 (old id 1760680)
- date added to LUP
- 2016-04-01 10:38:20
- date last changed
- 2022-03-12 07:37:28
@article{1fdd62b1-df73-485c-b248-01dd724c47b7, abstract = {{Protein S has an established role in the protein C anticoagulant pathway, where it enhances the factor Va (FVa) and factor VIIIa (FVIIIa) inactivating property of activated protein C (APC). Despite its physiological role and clinical importance, the molecular basis of its action is not fully understood. To clarify the mechanism of the protein S interaction with APC, we have constructed and expressed a library of composite or point variants of human protein S, with residue substitutions introduced into the Gla, thrombin-sensitive region (TSR), epidermal growth factor 1 (EGF1), and EGF2 domains. Cofactor activity for APC was evaluated by calibrated automated thrombography (CAT) using protein S-deficient plasma. Of 27 variants tested initially, only one, protein S D95A (within the EGF1 domain), was largely devoid of functional APC cofactor activity. Protein S D95A was, however, gamma-carboxylated and bound phospholipids with an apparent dissociation constant (Kd(app)) similar to that of wildtype (WT) protein S. In a purified assay using FVa R506Q/R679Q, purified protein S D95A was shown to have greatly reduced ability to enhance APC-induced cleavage of FVa Arg306. It is concluded that residue Asp95 within EGF1 is critical forAPC cofactor function of protein S and could define a principal functional interaction site for APC. (Blood. 2010;115(23):4878-4885)}}, author = {{Andersson, Helena M. and Arantes, Marcia J. and Crawley, James T. B. and Luken, Brenda M. and Tran, Sinh and Dahlbäck, Björn and Lane, David A. and Rezende, Suely M.}}, issn = {{1528-0020}}, language = {{eng}}, number = {{23}}, pages = {{4878--4885}}, publisher = {{American Society of Hematology}}, series = {{Blood}}, title = {{Activated protein C cofactor function of protein S: a critical role for Asp95 in the EGF1-like domain}}, url = {{http://dx.doi.org/10.1182/blood-2009-11-256610}}, doi = {{10.1182/blood-2009-11-256610}}, volume = {{115}}, year = {{2010}}, }