4C29 : Crystal Structure of High-Affinity von Willebrand Factor A1 domain with Disulfide Mutation

Creators

Mark A. Blenner, Dept. of Chemical and Biomolecular Engineering, Clemson University, Earle Hall
Xianchi Dong, Program in Cellular and Molecular Medicine, Boston Children's Hospital, Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School
Timothy A. Springer, Program in Cellular and Molecular Medicine, Boston Children's Hospital, Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School

Description

Experimental Technique/Method:X-RAY DIFFRACTION Resolution:2.2 Classification:BLOOD CLOTTING Release Date:2014-01-08 Deposition Date:2013-08-16 Revision Date:2014-01-15#2014-01-29#2014-03-12 Molecular Weight:50017.11 Macromolecule Type:Protein Residue Count:430 Atom Site Count:3269 DOI:10.2210/pdb4c29/pdb Abstract: Activation by elongational flow of von Willebrand factor (VWF) is critical for primary hemostasis. Mutations causing type 2B von Willebrand disease (VWD), platelet-type VWD (PT-VWD), and tensile force each increase affinity of the VWF A1 domain and platelet glycoprotein Ibα (GPIbα) for one another; however, the structural basis for these observations remains elusive. Directed evolution was used to discover a further gain-of-function mutation in A1 that shifts the long range disulfide bond by one residue. We solved multiple crystal structures of this mutant A1 and A1 containing two VWD mutations complexed with GPIbα containing two PT-VWD mutations. We observed a gained interaction between A1 and the central leucine-rich repeats (LRRs) of GPIbα, previously shown to be important at high shear stress, and verified its importance mutationally. These findings suggest that structural changes, including central GPIbα LRR-A1 contact, contribute to VWF affinity regulation. Among the mutant complexes, variation in contacts and poor complementarity between the GPIbα β-finger and the region of A1 harboring VWD mutations lead us to hypothesize that the structures are on a pathway to, but have not yet reached, a force-induced super high affinity state.

Publication Date

1-8-2014

Publisher

RCSB-PDB

DOI

10.2210/pdb4c29/pdb

Document Type

Data Set

Recommended Citation

Blenner, Mark A.; Dong, Xianchi; Springer, Timothy A. (2014), "4C29 : Crystal Structure of High-Affinity von Willebrand Factor A1 domain with Disulfide Mutation", RCSB-PDB, doi: 10.2210/pdb4c29/pdb

https://doi.org/10.2210/pdb4c29/pdb

Identifier

4C29

Embargo Date

1-8-2014