rFVIIIFc is composed of three polypeptide chains. Chain A is the heavy chain of FVIII. Chain B is the light chain of FVIII with a single human IgG1 Fc domain fused to the C terminus of the light chain. Chain C is a free single human IgG1 Fc domain that pairs with the Fc domain at the C terminus of the FVIII light chain. These three chains form the full assembly--a monomeric FVIII with a C-terminal, dimeric Fc-fusion. While all three chains are in the crystal, the dimeric Fc is not visible in the final structure. Silver stain gel of a dissolved crystal confirmed the presence of all three chains.
Authors state the following: The crystal contains a total of 3 polypeptide chains. Chain A is the ...Authors state the following: The crystal contains a total of 3 polypeptide chains. Chain A is the heavy chain of FVIII. Chain B is the light chain of FVIII fused to a single IgG1 Fc domain. The third chain, which is not indicated here, is another single IgG1 Fc domain that forms a dimer with the single Fc at the C-terminus of the FVIII light chain. There are no coordinates for the either of these Fc halves as there is no density for the dimeric Fc at the C-terminus of FVIII. We have dissolved crystals on a reducing gel and observe the presence of both halves of the Fc (one fused to the C terminus of the FVIII light chain and one free Fc). There are large solvent channels in the crystal that are large enough to accommodate the Fc dimer at the C-terminus of FVIII
Has protein modification
Y
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実験情報
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実験
実験
手法: X線回折 / 使用した結晶の数: 1
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試料調製
結晶
マシュー密度: 4.61 Å3/Da / 溶媒含有率: 73.33 %
結晶化
温度: 297 K / 手法: 蒸気拡散法 / pH: 7.3 / 詳細: 5% Ethanol, 100mM TRIS pH 7.3
解像度: 4.19→37.6 Å / Cor.coef. Fo:Fc: 0.851 / Cor.coef. Fo:Fc free: 0.715 / SU B: 146.864 / SU ML: 0.828 / 交差検証法: THROUGHOUT / σ(F): 0 / ESU R Free: 0.996 / 立体化学のターゲット値: MAXIMUM LIKELIHOOD 詳細: HYDROGENS HAVE BEEN USED IF PRESENT IN THE INPUT U VALUES : WITH TLS ADDED. Authors state the following: The crystal contains a total of 3 polypeptide chains. Chain A is the heavy chain of ...詳細: HYDROGENS HAVE BEEN USED IF PRESENT IN THE INPUT U VALUES : WITH TLS ADDED. Authors state the following: The crystal contains a total of 3 polypeptide chains. Chain A is the heavy chain of FVIII. Chain B is the light chain of FVIII fused to a single IgG1 Fc domain. The third chain, which is not indicated here, is another single IgG1 Fc domain that forms a dimer with the single Fc at the C-terminus of the FVIII light chain. There are no coordinates for the either of these Fc halves as there is no density for the dimeric Fc at the C-terminus of FVIII. We have dissolved crystals on a reducing gel and observe the presence of both halves of the Fc (one fused to the C terminus of the FVIII light chain and one free Fc). There are large solvent channels in the crystal that are large enough to accommodate the Fc dimer at the C-terminus of FVIII
Rfactor
反射数
%反射
Selection details
Rfree
0.3619
1309
5.1 %
RANDOM
Rwork
0.298
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obs
0.3011
24550
98.77 %
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溶媒の処理
イオンプローブ半径: 0.8 Å / 減衰半径: 0.8 Å / VDWプローブ半径: 1.2 Å / 溶媒モデル: MASK