PDBsum entry 1f7e

Blood clotting

PDB id: 1f7e

Contents

Protein chain

46 a.a. *

* Residue conservation analysis

PDB id:

1f7e

Name:

Blood clotting

Title:

The first egf-like domain from human blood coagulation fvii, nmr, 20 structures

Structure:

Protein (blood coagulation factor vii). Chain: a. Fragment: first egf-like domain (residues 45-87). Engineered: yes. Other_details: calcium bound form

Source:

Homo sapiens. Human. Organism_taxid: 9606. Expressed in: escherichia coli. Expression_system_taxid: 562

NMR struc:

20 models

Authors:

Y.-H.Kao,G.F.Lee,Y.Wang,M.A.Starovasnik,R.F.Kelley,M.W.Spellman, L.Lerner

Key ref:

Y.H.Kao et al. (1999). The effect of O-fucosylation on the first EGF-like domain from human blood coagulation factor VII. Biochemistry, 38, 7097-7110. PubMed id: 10353820 DOI: 10.1021/bi990234z

Date:

19-Feb-99 Release date: 16-Jun-99

Protein chain

P08709  (FA7_HUMAN) -  Coagulation factor VII from Homo sapiens

Seq: 466 a.a.

Struc: 46 a.a.*

* PDB and UniProt seqs differ at 3 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.3.4.21.21 - coagulation factor VIIa.
• Reaction: Hydrolyzes one Arg-|-Ile bond in factor X to form factor Xa.

DOI no: 10.1021/bi990234z

Biochemistry 38:7097-7110 (1999)

PubMed id: 10353820

The effect of O-fucosylation on the first EGF-like domain from human blood coagulation factor VII.

Y.H.Kao, G.F.Lee, Y.Wang, M.A.Starovasnik, R.F.Kelley, M.W.Spellman, L.Lerner.

ABSTRACT

The first epidermal growth factor-like domain (EGF-1) from blood coagulation factor VII (FVII) contains two unusual O-linked glycosylation sites at Ser-52 and Ser-60. We report here a detailed study of the effect of O-fucosylation at Ser-60 on the structure of FVII EGF-1, its Ca2+-binding affinity, and its interaction with tissue factor (TF). The in vitro fucosylation of the nonglycosylated FVII EGF-1 was achieved by using O-fucosyltransferase purified from Chinese hamster ovary cells. Distance and dihedral constraints derived from NMR data were used to determine the solution structures of both nonglycosylated and fucosylated FVII EGF-1 in the presence of CaCl2. The overall structure of fucosylated FVII EGF-1 is very similar to the nonfucosylated form even for the residues near the fucosylation site. The Ca2+ dissociation constants (Kd) for the nonfucosylated and fucosylated FVII EGF-1 were found to be 16.4 +/- 1.8 and 8.6 +/- 1.4 mM, respectively. The FVII EGF-1 domain binds to the extracellular part of TF with a low affinity (Kd approximately 0. 6 mM), and the addition of fucose appears to have no effect on this affinity. These results indicate that the FVII EGF-1 alone cannot form a tight complex with TF and suggest that the high binding affinity of FVIIa for TF requires cooperative interaction among the four domains in FVII with TF. Although the fucose has no significant effect on the interaction between TF and the individual FVII EGF-1 domain, it may affect the interaction of full-length FVIIa with TF by influencing its Ca2+-binding affinity.