Chun, Haarin Kurasawa, James H. Olivares, Philip Marakasova, Ekaterina S. Shestopal, Svetlana A. Hassink, Gabriela U. Karnaukhova, Elena Migliorini, Mary Obi, Juliet O. Smith, Ally K. Wintrode, Patrick L. Durai, Prasannavenkatesh Park, Keunwan Deredge, Daniel Strickland, Dudley K. Sarafanov, Andrey G. 2024-01-19T11:02:53Z 2024-01-19T11:02:53Z 2022-08-18 2022-10 1538-7933 https://pubs.kist.re.kr/handle/201004/114523 Background Deficiency in blood coagulation factor VIII (FVIII) results in life-threating bleeding (hemophilia A) treated by infusions of FVIII concentrates. To improve disease treatment, FVIII has been modified to increase its plasma half-life, which requires understanding mechanisms of FVIII catabolism. An important catabolic actor is hepatic low density lipoprotein receptor-related protein 1 (LRP1), which also regulates many other clinically significant processes. Previous studies showed complexity of FVIII site for binding LRP1. Objectives To characterize binding sites between FVIII and LRP1 and suggest a model of the interaction. Methods A series of recombinant ligand-binding complement-type repeat (CR) fragments of LRP1 including mutated variants was generated in a baculovirus system and tested for FVIII interaction using surface plasmon resonance, tissue culture model, hydrogen-deuterium exchange mass spectrometry, and in silico. Results Multiple CR doublets within LRP1 clusters II and IV were identified as alternative FVIII-binding sites. These interactions follow the canonical binding mode providing major binding energy, and additional weak interactions are contributed by adjacent CR domains. A representative CR doublet was shown to have multiple contact sites on FVIII. Conclusions FVIII and LRP1 interact via formation of multiple complex contacts involving both canonical and non-canonical binding combinations. We propose that FVIII-LRP1 interaction occurs via switching such alternative binding combinations in a dynamic mode, and that this mechanism is relevant to other ligand interactions of the low-density lipoprotein receptor family members including LRP1. English Blackwell Publishing Inc. Characterization of interaction between blood coagulation factor VIII and LRP1 suggests dynamic binding by alternating complex contacts Article 10.1111/jth.15817 1 Journal of Thrombosis and Haemostasis, v.20, no.10, pp.2255 - 2269 Journal of Thrombosis and Haemostasis 20 10 2255 2269 Y scie scopus 000836025300001 Hematology Peripheral Vascular Disease Hematology Cardiovascular System & Cardiology Article DENSITY-LIPOPROTEIN-RECEPTOR VON-WILLEBRAND-FACTOR CONSERVED ACIDIC RESIDUE HIGH-AFFINITY BINDING LIGAND-BINDING PROTEIN RAP BIVALENT COMPLEX FACTOR IXA RECOGNITION CLUSTER blood coagulation factor VIII hemophilia A LDL-receptor related protein-associated protein low density lipoprotein receptor low density lipoprotein receptor-related protein 1