PDBsum entry 1pfx

Hydrolase/hydrolase inhibitor

PDB id: 1pfx

Contents

Protein chains

235 a.a. *

146 a.a. *

Ligands

0G6

* Residue conservation analysis

PDB id:

1pfx

Name:

Hydrolase/hydrolase inhibitor

Title:

Porcine factor ixa

Structure:

Factor ixa. Chain: c. Synonym: christmas factor. Engineered: yes. Factor ixa. Chain: l. Synonym: christmas factor. Engineered: yes

Source:

Sus scrofa. Pig. Organism_taxid: 9823. Organ: blood. Gene: pete. Gene: pete

Biol. unit:

Dimer (from PQS)

Resolution:

3.00Å R-factor: 0.198

Authors:

H.Brandstetter,M.Bauer,R.Huber,P.Lollar,W.Bode

Key ref:

H.Brandstetter et al. (1995). X-ray structure of clotting factor IXa: active site and module structure related to Xase activity and hemophilia B. Proc Natl Acad Sci U S A, 92, 9796-9800. PubMed id: 7568220 DOI: 10.1073/pnas.92.21.9796

Date:

19-Jul-95 Release date: 17-Aug-96

Protein chain

P16293  (FA9_PIG) -  Coagulation factor IX (Fragment) from Sus scrofa

Seq: 409 a.a.

Struc: 235 a.a.*

Protein chain

P16293  (FA9_PIG) -  Coagulation factor IX (Fragment) from Sus scrofa

Seq: 409 a.a.

Struc: 146 a.a.*

* PDB and UniProt seqs differ at 15 residue positions (black crosses)

Enzyme reactions

• Enzyme class: Chains C, L: E.C.3.4.21.22 - coagulation factor IXa.
• Reaction: Hydrolyzes one Arg-|-Ile bond in factor X to form factor Xa.

DOI no: 10.1073/pnas.92.21.9796

Proc Natl Acad Sci U S A 92:9796-9800 (1995)

PubMed id: 7568220

X-ray structure of clotting factor IXa: active site and module structure related to Xase activity and hemophilia B.

H.Brandstetter, M.Bauer, R.Huber, P.Lollar, W.Bode.

ABSTRACT

Hereditary deficiency of factor IXa (fIXa), a key enzyme in blood coagulation, causes hemophilia B, a severe X chromosome-linked bleeding disorder afflicting 1 in 30,000 males; clinical studies have identified nearly 500 deleterious variants. The x-ray structure of porcine fIXa described here shows the atomic origins of the disease, while the spatial distribution of mutation sites suggests a structural model for factor X activation by phospholipid-bound fIXa and cofactor VIIIa. The 3.0-A-resolution diffraction data clearly show the structures of the serine proteinase module and the two preceding epidermal growth factor (EGF)-like modules; the N-terminal Gla module is partially disordered. The catalytic module, with covalent inhibitor D-Phe-1I-Pro-2I-Arg-3I chloromethyl ketone, most closely resembles fXa but differs significantly at several positions. Particularly noteworthy is the strained conformation of Glu-388, a residue strictly conserved in known fIXa sequences but conserved as Gly among other trypsin-like serine proteinases. Flexibility apparent in electron density together with modeling studies suggests that this may cause incomplete active site formation, even after zymogen, and hence the low catalytic activity of fIXa. The principal axes of the oblong EGF-like domains define an angle of 110 degrees, stabilized by a strictly conserved and fIX-specific interdomain salt bridge. The disorder of the Gla module, whose hydrophobic helix is apparent in electron density, can be attributed to the absence of calcium in the crystals; we have modeled the Gla module in its calcium form by using prothrombin fragment 1. The arched module arrangement agrees with fluorescence energy transfer experiments. Most hemophilic mutation sites of surface fIX residues occur on the concave surface of the bent molecule and suggest a plausible model for the membrane-bound ternary fIXa-FVIIIa-fX complex structure: fIXa and an equivalently arranged fX arch across an underlying fVIIIa subdomain from opposite sides; the stabilizing fVIIIa interactions force the catalytic modules together, completing fIXa active site formation and catalytic enhancement.