title: Factor VIII Interacts with the Endocytic Receptor Low-density Lipoprotein Receptor-related Protein 1 via an Extended Surface Comprising "Hot-Spot" Lysine Residues.
abstract: Lysine residues are implicated in driving the ligand binding to the LDL receptor family. However, it has remained unclear how specificity is regulated. Using coagulation factor VIII as a model ligand, we now study the contribution of individual lysine residues in the interaction with the largest member of the LDL receptor family, low-density lipoprotein receptor-related protein (LRP1). Using hydrogen-deuterium exchange mass spectrometry (HDX-MS) and SPR interaction analysis on a library of lysine replacement variants as two independent approaches, we demonstrate that the interaction between factor VIII (FVIII) and LRP1 occurs over an extended surface containing multiple lysine residues. None of the individual lysine residues account completely for LRP1 binding, suggesting an additive binding model. Together with structural docking studies, our data suggest that FVIII interacts with LRP1 via an extended surface of multiple lysine residues that starts at the bottom of the C1 domain and winds around the FVIII molecule.
authors.affiliation: From the Department of Plasma Proteins, Sanquin Blood Supply Foundation, 1066 CX Amsterdam, The Netherlands, m.vandenbiggelaar@sanquin.nl.
authors.forename: Maartje
authors.initials: M
authors.lastname: van den Biggelaar
authors.affiliation: Global Research, Novo Nordisk A/S, DK-2760 Måløv, Denmark, and.
authors.forename: Jesper J
authors.initials: JJ
authors.lastname: Madsen
authors.affiliation: Global Research, Novo Nordisk A/S, DK-2760 Måløv, Denmark, and.
authors.forename: Johan H
authors.initials: JH
authors.lastname: Faber
authors.affiliation: From the Department of Plasma Proteins, Sanquin Blood Supply Foundation, 1066 CX Amsterdam, The Netherlands.
authors.forename: Marleen G
authors.initials: MG
authors.lastname: Zuurveld
authors.affiliation: From the Department of Plasma Proteins, Sanquin Blood Supply Foundation, 1066 CX Amsterdam, The Netherlands.
authors.forename: Carmen
authors.initials: C
authors.lastname: van der Zwaan
authors.affiliation: Global Research, Novo Nordisk A/S, DK-2760 Måløv, Denmark, and.
authors.forename: Ole H
authors.initials: OH
authors.lastname: Olsen
authors.affiliation: Global Research, Novo Nordisk A/S, DK-2760 Måløv, Denmark, and.
authors.forename: Henning R
authors.initials: HR
authors.lastname: Stennicke
authors.affiliation: From the Department of Plasma Proteins, Sanquin Blood Supply Foundation, 1066 CX Amsterdam, The Netherlands, the Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, 3508 TC Utrecht, The Netherlands.
authors.forename: Koen
authors.initials: K
authors.lastname: Mertens
authors.affiliation: From the Department of Plasma Proteins, Sanquin Blood Supply Foundation, 1066 CX Amsterdam, The Netherlands, the Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, 3508 TC Utrecht, The Netherlands.
authors.forename: Alexander B
authors.initials: AB
authors.lastname: Meijer
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001665:Binding Sites; D018360:Crystallography, X-Ray; D041961:Deuterium Exchange Measurement; D004705:Endocytosis; D005169:Factor VIII / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008077:Lipoproteins, LDL / Q000378:metabolism; D026503:Low Density Lipoprotein Receptor-Related Protein-1 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008239:Lysine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary
chemical_list: C475476:LRP1 protein, human; D008077:Lipoproteins, LDL; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D005169:Factor VIII; D008239:Lysine
keywords: coagulation factor; factor VIII (FVIII); hydrogen exchange mass spectrometry; hydrogen-deuterium exchange; low-density lipoprotein receptor-related protein 1; protein-protein interaction; receptor; surface plasmon resonance (SPR)