Stoichiometry and architecture of the platelet membrane complex glycoprotein Ib-IX-V
Author:
Lu Juanjuan12, Zhang Chunli12, Shi Shaohua12, Li Shaobai12, Liu Junling3, Wu Jian12, Huang Chenhui12, Lei Ming124
Affiliation:
1. Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine , Shanghai , China 2. Shanghai Institute of Precision Medicine , Shanghai , China 3. Department of Biochemistry and Molecular Cell Biology , Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Shanghai Jiao Tong University School of Medicine , Shanghai , China 4. State Key laboratory of Oncogenes and Related Genes, Shanghai Jiao Tong University School of Medicine , Shanghai , China
Abstract
Abstract
Glycoprotein (GP) Ib-IX-V is the second most abundant platelet receptor for thrombin and other ligands crucial for hemostasis and thrombosis. Its activity is involved in platelet adhesion to vascular injury sites and thrombin-induced platelet aggregation. GPIb-IX-V is a heteromeric complex composed of four subunits, GPIbα, GPIbβ, GPV and GPIX, in a stoichiometric ratio that has been wildly debated. Despite its important physiological roles, the overall structure and molecular arrangement of GPIb-IX-V are not yet fully understood. Here, we purify stable and functional human GPIb-IX-V complex from reconstituted EXPi293F cells in high homogeneity, and perform biochemical and structural characterization of this complex. Single-particle cryo-electron microscopy structure of GPIb-IX-V is determined at ∼11 Å resolution, which unveils the architecture of GPIb-IX-V and its subunit organization. Size-exclusion chromatography-multi-angle static light scattering analysis reveals that GPIb-IX-V contains GPIb-IX and GPV at a 1:1 stoichiometric ratio and surface plasmon resonance assays show that association of GPV leads to slow kinetics of thrombin binding to GPIb-IX-V. Taken together, our results provide the first three-dimensional architecture of the intact GPIb-IX-V complex, which extends our understanding of the structure and functional mechanism of this complex in hemostasis and thrombosis.
Funder
National Key Research and Development Program of China National Key R&D Program of China Fundamental research program funding of Ninth People’s Hospital affiliated to Shanghai Jiao Tong University School of Medicine
Publisher
Walter de Gruyter GmbH
Subject
Clinical Biochemistry,Molecular Biology,Biochemistry
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