Table 2.

FIXa regions mediating the interaction with FVIIIa

FIXa domain or regionResiduesFVIIIa counterpartExperimental strategy and references
Legacy numberingChymotrypsinogen numberingDomainLegacy numbering
Gla  Y1-S3
E21-F25
V46-D47 
n.a. C2 E2228-V2240 Gla peptides41
rFVIII domains/peptides43
Small-angle X-ray scattering33  
Gla§  N34-D47 n.a. C1 S2113-N2118 Computational docking and MD simulations32  
EGF1§  N67, S68
E78, and K80 
n.a. A3 T1695, H1697
T1739, and Q1745-Y1748 
Computational docking and MD simulations32  
EGF2 T87-D104 n.a. A3 Q1692-H1697
N1770-M1772
E1793-Q1798
T1811-K1818 
rFIX variants38,42
rFVIII variants44,46
Computational docking and MD simulations32  
Protease, 99c-loop Y259-Y266 Y94-Y99 A3 I1790-Q1798
R1803-K1818 
rFVIII variants44,46
Small-angle X-ray scattering33  
Protease, exosite II K293-K301 K126-K132 A2, A3 R698, K707-N714
E1793, Q1796 
rFVIII variants45,46
HDX-MS of rFIX variants27
Computational docking and MD simulations32  
Protease, exosite II L330-R338 L162-R170 A2 S558-M567 rFIX variants; rFVIII domains/peptides39
HDX-MS of rFIX variants27
Computational docking and MD simulations32  
Protease, exosite II N406-E410 N236-E240 a2 E738 HDX-MS of rFIX variants27
Computational docking and MD simulations32  
FIXa domain or regionResiduesFVIIIa counterpartExperimental strategy and references
Legacy numberingChymotrypsinogen numberingDomainLegacy numbering
Gla  Y1-S3
E21-F25
V46-D47 
n.a. C2 E2228-V2240 Gla peptides41
rFVIII domains/peptides43
Small-angle X-ray scattering33  
Gla§  N34-D47 n.a. C1 S2113-N2118 Computational docking and MD simulations32  
EGF1§  N67, S68
E78, and K80 
n.a. A3 T1695, H1697
T1739, and Q1745-Y1748 
Computational docking and MD simulations32  
EGF2 T87-D104 n.a. A3 Q1692-H1697
N1770-M1772
E1793-Q1798
T1811-K1818 
rFIX variants38,42
rFVIII variants44,46
Computational docking and MD simulations32  
Protease, 99c-loop Y259-Y266 Y94-Y99 A3 I1790-Q1798
R1803-K1818 
rFVIII variants44,46
Small-angle X-ray scattering33  
Protease, exosite II K293-K301 K126-K132 A2, A3 R698, K707-N714
E1793, Q1796 
rFVIII variants45,46
HDX-MS of rFIX variants27
Computational docking and MD simulations32  
Protease, exosite II L330-R338 L162-R170 A2 S558-M567 rFIX variants; rFVIII domains/peptides39
HDX-MS of rFIX variants27
Computational docking and MD simulations32  
Protease, exosite II N406-E410 N236-E240 a2 E738 HDX-MS of rFIX variants27
Computational docking and MD simulations32  

HDX-MS, hydrogen-deuterium exchange-mass spectrometry; MD, molecular dynamics; n.a., not applicable; r, recombinant.

Legacy numbering represents mature FIX with residue 1 corresponding to the first residue of the Gla domain (which equals residue 47 in the numbering format of the Human Genome Variation Society).132 

Chymotrypsinogen numbering of the serine protease domain, as originally described by Bode et al13 

FIXa-FVIIIa model in which the Gla domain is assumed to interact with the C2 domain of FVIIIa.

§

FIXa-FVIIIa model in which the Gla domain is assumed to interact with the C1 domain of FVIIIa.

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