C-terminal residues of activated protein C light chain contribute to its anticoagulant and cytoprotective activities.
activated protein C
anticoagulant activity
cytoprotective activity
protease activated receptor
protein S
Journal
Journal of thrombosis and haemostasis : JTH
ISSN: 1538-7836
Titre abrégé: J Thromb Haemost
Pays: England
ID NLM: 101170508
Informations de publication
Date de publication:
05 2020
05 2020
Historique:
received:
14
08
2019
revised:
23
01
2020
accepted:
28
01
2020
pubmed:
6
2
2020
medline:
15
5
2021
entrez:
5
2
2020
Statut:
ppublish
Résumé
Activated protein C (APC) is an important homeostatic blood coagulation protease that conveys anticoagulant and cytoprotective activities. Proteolytic inactivation of factors Va and VIIIa facilitated by cofactor protein S is responsible for APC's anticoagulant effects, whereas cytoprotective effects of APC involve primarily the endothelial protein C receptor (EPCR), protease activated receptor (PAR)1 and PAR3. To date, several binding exosites in the protease domain of APC have been identified that contribute to APC's interaction with its substrates but potential contributions of the C-terminus of the light chain have not been studied in detail. Site-directed Ala-scanning mutagenesis of six positively charged residues within G142-L155 was used to characterize their contributions to APC's anticoagulant and cytoprotective activities. K151 was involved in protein S dependent-anticoagulant activity of APC with some contribution of K150. 3D structural analysis supported that these two residues were exposed in an extended protein S binding site on one face of APC. Both K150 and K151 were important for PAR1 and PAR3 cleavage by APC, suggesting that this region may also mediate interactions with PARs. Accordingly, APC's cytoprotective activity as determined by endothelial barrier protection was impaired by Ala substitutions of these residues. Thus, both K150 and K151 are involved in APC's anticoagulant and cytoprotective activities. The differential contribution of K150 relative to K151 for protein S-dependent anticoagulant activity and PAR cleavage highlights that binding exosites for protein S binding and for PAR cleavage in the C-terminal region of APC's light chain overlap.
Sections du résumé
BACKGROUND
Activated protein C (APC) is an important homeostatic blood coagulation protease that conveys anticoagulant and cytoprotective activities. Proteolytic inactivation of factors Va and VIIIa facilitated by cofactor protein S is responsible for APC's anticoagulant effects, whereas cytoprotective effects of APC involve primarily the endothelial protein C receptor (EPCR), protease activated receptor (PAR)1 and PAR3.
OBJECTIVE
To date, several binding exosites in the protease domain of APC have been identified that contribute to APC's interaction with its substrates but potential contributions of the C-terminus of the light chain have not been studied in detail.
METHODS
Site-directed Ala-scanning mutagenesis of six positively charged residues within G142-L155 was used to characterize their contributions to APC's anticoagulant and cytoprotective activities.
RESULTS AND CONCLUSIONS
K151 was involved in protein S dependent-anticoagulant activity of APC with some contribution of K150. 3D structural analysis supported that these two residues were exposed in an extended protein S binding site on one face of APC. Both K150 and K151 were important for PAR1 and PAR3 cleavage by APC, suggesting that this region may also mediate interactions with PARs. Accordingly, APC's cytoprotective activity as determined by endothelial barrier protection was impaired by Ala substitutions of these residues. Thus, both K150 and K151 are involved in APC's anticoagulant and cytoprotective activities. The differential contribution of K150 relative to K151 for protein S-dependent anticoagulant activity and PAR cleavage highlights that binding exosites for protein S binding and for PAR cleavage in the C-terminal region of APC's light chain overlap.
Identifiants
pubmed: 32017367
doi: 10.1111/jth.14756
pmc: PMC7380734
mid: NIHMS1553564
pii: S1538-7836(22)00307-5
doi:
Substances chimiques
Anticoagulants
0
Protein C
0
Receptor, PAR-1
0
Factor Va
65522-14-7
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1027-1038Subventions
Organisme : NHLBI NIH HHS
ID : R01 HL142975
Pays : United States
Organisme : NHLBI NIH HHS
ID : R37 HL052246
Pays : United States
Organisme : NIGMS NIH HHS
ID : GM09888
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL104165
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL052246
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL130678
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL148096
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM096888
Pays : United States
Informations de copyright
© 2020 International Society on Thrombosis and Haemostasis.
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