The roles of factor Va and protein S in formation of the activated protein C/protein S/factor Va inactivation complex.
Binding Sites
Blood Coagulation
Calcium-Binding Proteins
/ chemistry
Enzyme Activation
Factor Va
/ chemistry
HEK293 Cells
Humans
Models, Molecular
Multiprotein Complexes
Phospholipids
/ chemistry
Protein Binding
Protein C
/ chemistry
Protein Conformation
Protein S
/ chemistry
Structure-Activity Relationship
Thrombin
/ metabolism
Thromboplastin
/ metabolism
activated protein C
factor Va
phopsholipids
protein S
prothrombinase
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:
12 2019
12 2019
Historique:
received:
06
03
2019
accepted:
26
07
2019
pubmed:
1
8
2019
medline:
9
9
2020
entrez:
1
8
2019
Statut:
ppublish
Résumé
Activated protein C (APC)-mediated inactivation of factor (F)Va is greatly enhanced by protein S. For inactivation to occur, a trimolecular complex among FVa, APC, and protein S must form on the phospholipid membrane. However, direct demonstration of complex formation has proven elusive. To elucidate the nature of the phospholipid-dependent interactions among APC, protein S, and FVa. We evaluated binding of active site blocked APC to phospholipid-coated magnetic beads in the presence and absence of protein S and/or FVa. The importance of protein S and FV residues were evaluated functionally. Activated protein C alone bound weakly to phospholipids. Protein S mildly enhanced APC binding to phospholipid surfaces, whereas FVa did not. However, FVa together with protein S enhanced APC binding (>14-fold), demonstrating formation of an APC/protein S/FVa complex. C4b binding protein-bound protein S failed to enhance APC binding, agreeing with its reduced APC cofactor function. Protein S variants (E36A and D95A) with reduced APC cofactor function exhibited essentially normal augmentation of APC binding to phospholipids, but diminished APC/protein S/FVa complex formation, suggesting involvement in interactions dependent upon FVa. Similarly, FVa FVa plays a central role in the formation of its inactivation complex. Furthermore, membrane proximal interactions among FVa, APC, and protein S are essential for its cofactor function.
Sections du résumé
BACKGROUND
Activated protein C (APC)-mediated inactivation of factor (F)Va is greatly enhanced by protein S. For inactivation to occur, a trimolecular complex among FVa, APC, and protein S must form on the phospholipid membrane. However, direct demonstration of complex formation has proven elusive.
OBJECTIVES
To elucidate the nature of the phospholipid-dependent interactions among APC, protein S, and FVa.
METHODS
We evaluated binding of active site blocked APC to phospholipid-coated magnetic beads in the presence and absence of protein S and/or FVa. The importance of protein S and FV residues were evaluated functionally.
RESULTS
Activated protein C alone bound weakly to phospholipids. Protein S mildly enhanced APC binding to phospholipid surfaces, whereas FVa did not. However, FVa together with protein S enhanced APC binding (>14-fold), demonstrating formation of an APC/protein S/FVa complex. C4b binding protein-bound protein S failed to enhance APC binding, agreeing with its reduced APC cofactor function. Protein S variants (E36A and D95A) with reduced APC cofactor function exhibited essentially normal augmentation of APC binding to phospholipids, but diminished APC/protein S/FVa complex formation, suggesting involvement in interactions dependent upon FVa. Similarly, FVa
CONCLUSIONS
FVa plays a central role in the formation of its inactivation complex. Furthermore, membrane proximal interactions among FVa, APC, and protein S are essential for its cofactor function.
Identifiants
pubmed: 31364267
doi: 10.1111/jth.14594
pmc: PMC6916587
pii: S1538-7836(22)03081-1
doi:
Substances chimiques
Calcium-Binding Proteins
0
Multiprotein Complexes
0
PROS1 protein, human
0
Phospholipids
0
Protein C
0
Protein S
0
Factor Va
65522-14-7
Thromboplastin
9035-58-9
Thrombin
EC 3.4.21.5
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2056-2068Subventions
Organisme : British Heart Foundation
ID : PG/14/90/31219
Pays : United Kingdom
Organisme : British Heart Foundation
ID : PG/14/63/31036
Pays : United Kingdom
Organisme : British Heart Foundation
ID : FS/10/004/28165
Pays : United Kingdom
Organisme : British Heart Foundation
ID : PG/17/42/33039
Pays : United Kingdom
Organisme : British Heart Foundation
ID : FS/12/60/29874
Pays : United Kingdom
Organisme : British Heart Foundation
ID : RG/18/3/33405
Pays : United Kingdom
Informations de copyright
© 2019 The Authors. Journal of Thrombosis and Haemostasis published by Wiley Periodicals, Inc. on behalf of International Society on Thrombosis and Haemostasis.
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