1FF7

THE FIRST EGF-LIKE DOMAIN FROM HUMAN BLOOD COAGULATION FVII (FUCOSYLATED AT SER-60), NMR, 20 STRUCTURES

Summary for 1FF7

• Entry DOI: 10.2210/pdb1ff7/pdb
• Descriptor: PROTEIN (Blood Coagulation Factor VII), alpha-L-fucopyranose (2 entities in total)
• Functional Keywords: factor vii, blood coagulation, egf-like domain, glycoprotein, fucosylation, o- linked fucose, blood clotting
• Biological source: Homo sapiens (human)
• Total number of polymer chains: 1
• Total formula weight: 5042.40

Authors

Kao, Y.-H.,Lee, G.F.,Wang, Y.,Starovasnik, M.A.,Kelley, R.F.,Spellman, M.W.,Lerner, L. (deposition date: 1999-02-19, release date: 1999-06-16, Last modification date: 2024-10-16)

Primary citation

Kao, Y.H.,Lee, G.F.,Wang, Y.,Starovasnik, M.A.,Kelley, R.F.,Spellman, M.W.,Lerner, L.
The effect of O-fucosylation on the first EGF-like domain from human blood coagulation factor VII.
Biochemistry, 38:7097-7110, 1999

PubMed 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.

PubMed: 10353820
DOI: 10.1021/bi990234z

Experimental method

SOLUTION NMR