Extensive small-angle X-ray scattering studies of blood coagulation factor VIIa reveal interdomain flexibility
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Extensive small-angle X-ray scattering studies of blood coagulation factor VIIa reveal interdomain flexibility. / Mosbæk, Charlotte Rode; Nolan, David; Persson, Egon; Svergun, Dmitri I; Bukrinsky, Jens Thostrup; Vestergaard, Bente.
In: Biochemistry, Vol. 49, No. 45, 2010, p. 9739-9745.Research output: Contribution to journal › Journal article › Research › peer-review
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T1 - Extensive small-angle X-ray scattering studies of blood coagulation factor VIIa reveal interdomain flexibility
AU - Mosbæk, Charlotte Rode
AU - Nolan, David
AU - Persson, Egon
AU - Svergun, Dmitri I
AU - Bukrinsky, Jens Thostrup
AU - Vestergaard, Bente
PY - 2010
Y1 - 2010
N2 - Blood coagulation factor VIIa (FVIIa) is used in the treatment of replacement therapy resistant hemophilia patients, and FVIIa is normally activated upon complex formation with tissue factor (TF), potentially in context with structural rearrangements. The solution behavior of uncomplexed FVIIa is important for understanding the mechanism of activation and for the stability and activity of the pharmaceutical product. However, crystal structures of FVIIa in complex with TF and of truncated free FVIIa reveal different overall conformations while previous small-angle scattering studies suggest FVIIa always to be fully extended in solution. Here, small-angle X-ray scattering analysis of multiple forms of FVIIa and TF under several experimental conditions elaborate extensively on the understanding of the solution behavior of FVIIa. We reveal significant FVIIa domain flexibility in solution, whereas TF has a well-defined conformation. Unspecific formation of dimers of FVIIa is also observed and varies with experimental conditions. In particular, active site-inhibited FVIIa displays a distinct solution behavior different from that of uninhibited FVIIa, which may reflect structural rearrangements causing resistance to activation, thereby emphasizing the connection between the distribution of different conformations of FVII and the mechanism of activation.
AB - Blood coagulation factor VIIa (FVIIa) is used in the treatment of replacement therapy resistant hemophilia patients, and FVIIa is normally activated upon complex formation with tissue factor (TF), potentially in context with structural rearrangements. The solution behavior of uncomplexed FVIIa is important for understanding the mechanism of activation and for the stability and activity of the pharmaceutical product. However, crystal structures of FVIIa in complex with TF and of truncated free FVIIa reveal different overall conformations while previous small-angle scattering studies suggest FVIIa always to be fully extended in solution. Here, small-angle X-ray scattering analysis of multiple forms of FVIIa and TF under several experimental conditions elaborate extensively on the understanding of the solution behavior of FVIIa. We reveal significant FVIIa domain flexibility in solution, whereas TF has a well-defined conformation. Unspecific formation of dimers of FVIIa is also observed and varies with experimental conditions. In particular, active site-inhibited FVIIa displays a distinct solution behavior different from that of uninhibited FVIIa, which may reflect structural rearrangements causing resistance to activation, thereby emphasizing the connection between the distribution of different conformations of FVII and the mechanism of activation.
KW - Former Faculty of Pharmaceutical Sciences
U2 - 10.1021/bi1011207
DO - 10.1021/bi1011207
M3 - Journal article
C2 - 20873866
VL - 49
SP - 9739
EP - 9745
JO - Biochemistry
JF - Biochemistry
SN - 0006-2960
IS - 45
ER -
ID: 23159787