The B domain of coagulation factor VIII interacts with the asialoglycoprotein receptor.
BACKGROUND
Coagulation factor VIII (FVIII) is a heavily glycosylated heterodimeric plasma protein that consists of a heavy (domains A1-A2-B) and light chain (domains A3-C1-C2). It has been well established that the clearance of FVIII from the circulation involves mechanisms that are sensitive to the low-density lipoprotein receptor (LDLR) family antagonist receptor-associated protein (RAP), including LDLR-related protein. Because FVIII clearance in the presence of a bolus injection of RAP still occurs fairly efficient, also RAP-independent mechanisms are likely to be involved.


OBJECTIVE
In the present study, we investigated the interaction of FVIII with the endocytic lectin asialoglycoprotein receptor (ASGPR) and the physiological relevance thereof.


RESULTS
Surface plasmon resonance studies demonstrated that FVIII dose-dependently bound to ASGPR with high affinity (Kd approximately 2 nM). FVIII subunits were different in that only the heavy chain displayed high-affinity binding to ASGPR. Studies employing a FVIII variant that lacks the B domain revealed that FVIII-ASGPR complex assembly is driven by structure elements within the B domain of the heavy chain. The FVIII heavy chain-ASGPR interaction required calcium ions and was inhibited by soluble D-galactose. Furthermore, deglycosylation of the FVIII heavy chain by endoglycosidase F completely abrogated the interaction with ASGPR. In clearance experiments in mice, the FVIII mean residence time was prolonged by the ASGPR-antagonist asialo-orosomucoid (ASOR).


CONCLUSIONS
We conclude that asparagine-linked oligosaccharide structures of the FVIII B domain recognize the carbohydrate recognition domains of ASGPR and that an ASOR-sensitive mechanism, most likely ASGPR, contributes to the catabolism of coagulation FVIII in vivo.
Bovenschen
N
N
Department of Plasma Proteins, Sanquin Research at CLB, Amsterdam, The Netherlands.
Rijken
D C
DC
Havekes
L M
LM
van Vlijmen
B J M
BJ
Mertens
K
K
D000818:Animals; D037263:Asialoglycoprotein Receptor / Q000378:metabolism*; D001665:Binding Sites; D002118:Calcium / Q000494:pharmacology; D002241:Carbohydrates; D005169:Factor VIII / Q000737:chemistry / Q000378:metabolism* / Q000493:pharmacokinetics; D005690:Galactose / Q000494:pharmacology; D006031:Glycosylation; D006801:Humans; D008297:Male; D008657:Metabolic Clearance Rate; D051379:Mice; D008810:Mice, Inbred C57BL; D011485:Protein Binding / Q000187:drug effects; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000378:metabolism; D020349:Surface Plasmon Resonance
D037263:Asialoglycoprotein Receptor; D002241:Carbohydrates; D021122:Protein Subunits; D005169:Factor VIII; D002118:Calcium; D005690:Galactose