Biosynthesis, assembly and secretion of coagulation factor VIII.

Article Details

Citation

Kaufman RJ, Pipe SW, Tagliavacca L, Swaroop M, Moussalli M

Biosynthesis, assembly and secretion of coagulation factor VIII.

Blood Coagul Fibrinolysis. 1997 Dec;8 Suppl 2:S3-14.

PubMed ID
9607108 [ View in PubMed
]
Abstract

Factor VIII is a large complex glycoprotein that is deficient in hemophilia A. It has a domain organization consisting of A1-A2-B-A3-C1-C2 where the B domain is a heavily glycosylated region that is dispensable for procoagulant activity. Factor VIII expression is 10-to 20-fold lower than the homologous coagulation factor V. Factor VIII expression is limited due to a low level of steady-state messenger RNA in the cytoplasm and inefficient transport of the primary translation product from the endoplasmic reticulum to the Golgi apparatus. Within the secretory pathway, factor VIII is processed to a heterodimer of the heavy chain (domains A1-A2-B) in a metal ion association with the light chain (domains A3-C1-C2). Upon secretion from the cell, von Willebrand factor binds the light chain of factor VIII and stabilizes the factor, preventing degradation. Protein folding within the mammalian secretory pathway is facilitated by molecular chaperones. Within the endoplasmic reticulum, factor VIII exhibits stable interaction with protein chaperones identified as the immunoglobulin-binding protein (BiP), calnexin and calreticulin. BiP is a peptide-dependent ATPase that interacts with exposed hydrophobic surfaces on unfolded proteins or unassembled protein subunits. A potential BiP binding site within factor VIII has been identified. Mutation of a single amino acid residue in the potential BiP binding site increased the secretion efficiency of factor VIII by threefold. Interestingly, the proposed BiP binding site is adjacent to a type-1 copper binding site within the A1 domain that is required for interaction between the factor VIII A1 domain and the A3 domain. We propose that Cu(I) binds the type-1 copper ion-binding site in the A1 domain and provides the essential requirement for a stable interaction between the heavy and light chains. Calnexin and calreticulin are transmembrane and lumenal proteins, respectively, localized to the endoplasmic reticulum, which associate transiently with many soluble and membrane glycoproteins during folding and subunit assembly. The calnexin and calreticulin interaction with factor VIII occurs primarily through amino-terminal linked oligosaccharides within the heavily glycosylated factor VIII B domain and this interaction appears to be required for factor VIII secretion. The findings suggest that factor VIII cycles through interactions with BiP, calnexin and calreticulin. Although the interaction with BiP does not appear to be required for factor VIII secretion, data suggest that the calnexin and/or calreticulin interaction is required for secretion. The observations suggest a unique requirement for carbohydrate processing and calnexin/calreticulin interaction that may limit the productive secretion of factor VIII and have implications for approaches towards somatic cell gene therapy for hemophilia A.

DrugBank Data that Cites this Article

Drug Targets
DrugTargetKindOrganismPharmacological ActionActions
Antihemophilic factor, human recombinant78 kDa glucose-regulated proteinProteinHumans
Unknown
Chaperone
Details
Antihemophilic factor, human recombinantCalnexinProteinHumans
Unknown
Chaperone
Details
Antihemophilic factor, human recombinantCalreticulinProteinHumans
Unknown
Chaperone
Details
Lonoctocog alfa78 kDa glucose-regulated proteinProteinHumans
Unknown
Chaperone
Details
Lonoctocog alfaCalnexinProteinHumans
Unknown
Chaperone
Details
Lonoctocog alfaCalreticulinProteinHumans
Unknown
Chaperone
Details
Moroctocog alfa78 kDa glucose-regulated proteinProteinHumans
Unknown
Chaperone
Details
Moroctocog alfaCalnexinProteinHumans
Unknown
Chaperone
Details
Moroctocog alfaCalreticulinProteinHumans
Unknown
Chaperone
Details