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PMID: 3262057 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

The role of beta-hydroxyaspartate and adjacent carboxylate residues in the first EGF domain of human factor IX.

The EMBO journal ·Vol. 7 ·No. 7 ·1988-07-00 ·Pages 2053-61

Rees DJ, Jones IM, Handford PA, Walter SJ, Esnouf MP, Smith KJ, Brownlee GG

Abstract

beta-Hydroxyaspartic acid is a post-translationally modified amino acid found in a number of plasma proteins in a domain homologous to epidermal growth factor. Its presence can be correlated with a high affinity Ca2+ binding site, with a dissociation constant of 10-100 microM. We describe a system for the expression of human coagulation factor IX in dog kidney cells in tissue culture, in which the post-translational modifications and the biochemical activity are indistinguishable from factor IX synthesized in vivo. This system has been used to express eight different point mutations of human factor IX in the first epidermal growth factor domain in order to study the role of beta-hydroxyaspartate at residue 64, and the adjacent carboxylate residues at positions 47, 49 and 78. We conclude that this domain is essential for factor IX function and suggest that Ca2+ binds to carboxylate ions in this domain and stabilizes a conformation necessary for the interaction of factor IXa with factor X, factor VIII and phospholipid in the next step of the clotting cascade.

MeSH Terms
Amino Acid Sequence Animals Aspartic Acid/analogs & derivatives,metabolism Carboxylic Acids/metabolism Codon Dogs Epidermal Growth Factor/genetics Factor IX/genetics,isolation & purification,metabolism Factor IXa Humans Macromolecular Substances Molecular Sequence Data Mutation Protein Processing, Post-Translational Serine Endopeptidases/metabolism
Chemicals
Carboxylic Acids Codon Macromolecular Substances 3-hydroxyaspartic acid Aspartic Acid Epidermal Growth Factor Factor IX Serine Endopeptidases Factor IXa
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Rees D J
Sir William Dunn School of Pathology, University of Oxford, UK.
Jones I M
Handford P A
Walter S J
Esnouf M P
Smith K J
Brownlee G G
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1988-07-00
Pages
2053-61
Language
English
Region
England
NLM ID
8208664
PMCID
PMC454485
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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