Abstract
Hemorrhagic factor V inhibitors frequently bind to the second C-type (C2) domain of factor V and interfere with phospholipid binding. To define specific residues recognized by inhibitors from four patients (one bovine thrombin-induced and three spontaneous antibodies), epitope mapping was performed using recombinant human factor V lacking most of the B-type domain (FV des B) and alanine-substituted mutants within the C2 domain (FV des B C2 mutants). FV des B C2 mutants located in the region between Lys2060 and Glu2069 were resistant to inhibition by three IgG preparations including the bovine thrombin-induced antibody in both prothrombinase and phospholipid-binding assays. In contrast, mutations at Lys2087 and Lys2092/Glu2096 were significantly resistant to inhibition by the fourth IgG preparation in both prothrombinase and phospholipid-binding assays. These results confirm interference of phospholipid binding by hemorrhagic factor V inhibitors and support the role(s) of these residues in phospholipid binding.
| Original language | English |
|---|---|
| Pages (from-to) | 1048-1054 |
| Number of pages | 7 |
| Journal | Thrombosis and Haemostasis |
| Volume | 85 |
| Issue number | 6 |
| Publication status | Published - 2001 Jul 5 |
| Externally published | Yes |
Fingerprint
Keywords
- C2 domain
- Epitope mapping
- Factor V
- Hemorrhagic inhibitor
- Phospholipid binding
ASJC Scopus subject areas
- Hematology
Cite this
Fine mapping of inhibitory anti-factor V antibodies using factor V C2 domain mutants : Identification of two antigenic epitopes involved in phospholipid binding. / Izumi, T.; Kim, Suhng Wook; Greist, A.; Macedo-Ribeiro, S.; Fuentes-Prior, P.; Bode, W.; Kane, W. H.; Ortel, T. L.
In: Thrombosis and Haemostasis, Vol. 85, No. 6, 05.07.2001, p. 1048-1054.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Fine mapping of inhibitory anti-factor V antibodies using factor V C2 domain mutants
T2 - Identification of two antigenic epitopes involved in phospholipid binding
AU - Izumi, T.
AU - Kim, Suhng Wook
AU - Greist, A.
AU - Macedo-Ribeiro, S.
AU - Fuentes-Prior, P.
AU - Bode, W.
AU - Kane, W. H.
AU - Ortel, T. L.
PY - 2001/7/5
Y1 - 2001/7/5
N2 - Hemorrhagic factor V inhibitors frequently bind to the second C-type (C2) domain of factor V and interfere with phospholipid binding. To define specific residues recognized by inhibitors from four patients (one bovine thrombin-induced and three spontaneous antibodies), epitope mapping was performed using recombinant human factor V lacking most of the B-type domain (FV des B) and alanine-substituted mutants within the C2 domain (FV des B C2 mutants). FV des B C2 mutants located in the region between Lys2060 and Glu2069 were resistant to inhibition by three IgG preparations including the bovine thrombin-induced antibody in both prothrombinase and phospholipid-binding assays. In contrast, mutations at Lys2087 and Lys2092/Glu2096 were significantly resistant to inhibition by the fourth IgG preparation in both prothrombinase and phospholipid-binding assays. These results confirm interference of phospholipid binding by hemorrhagic factor V inhibitors and support the role(s) of these residues in phospholipid binding.
AB - Hemorrhagic factor V inhibitors frequently bind to the second C-type (C2) domain of factor V and interfere with phospholipid binding. To define specific residues recognized by inhibitors from four patients (one bovine thrombin-induced and three spontaneous antibodies), epitope mapping was performed using recombinant human factor V lacking most of the B-type domain (FV des B) and alanine-substituted mutants within the C2 domain (FV des B C2 mutants). FV des B C2 mutants located in the region between Lys2060 and Glu2069 were resistant to inhibition by three IgG preparations including the bovine thrombin-induced antibody in both prothrombinase and phospholipid-binding assays. In contrast, mutations at Lys2087 and Lys2092/Glu2096 were significantly resistant to inhibition by the fourth IgG preparation in both prothrombinase and phospholipid-binding assays. These results confirm interference of phospholipid binding by hemorrhagic factor V inhibitors and support the role(s) of these residues in phospholipid binding.
KW - C2 domain
KW - Epitope mapping
KW - Factor V
KW - Hemorrhagic inhibitor
KW - Phospholipid binding
UR - http://www.scopus.com/inward/record.url?scp=0034970408&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034970408&partnerID=8YFLogxK
M3 - Article
C2 - 11434683
AN - SCOPUS:0034970408
VL - 85
SP - 1048
EP - 1054
JO - Thrombosis and Haemostasis
JF - Thrombosis and Haemostasis
SN - 0340-6245
IS - 6
ER -