PDBsum entry 4c29

Blood clotting

PDB id: 4c29

Contents

Protein chains

201 a.a.

Ligands

ACT

PEG ×4

Metals

_CA ×3

Waters ×175

PDB id:

4c29

Name:

Blood clotting

Title:

Crystal structure of high-affinity von willebrand factor a1 domain with disulfide mutation

Structure:

Von willebrand factor. Chain: a, b. Fragment: residues 1264-1471. Synonym: vwf, von willebrand antigen 2, von willebrand antigen ii. Engineered: yes. Mutation: yes. Other_details: high affinity mutant of a1

Source:

Homo sapiens. Human. Organism_taxid: 9606. Expressed in: escherichia coli. Expression_system_taxid: 469008.

Resolution:

2.20Å R-factor: 0.188 R-free: 0.235

Authors:

M.A.Blenner,X.Dong,T.A.Springer

Key ref:

M.A.Blenner et al. (2014). Structural basis of regulation of von Willebrand factor binding to glycoprotein Ib. J Biol Chem, 289, 5565-5579. PubMed id: 24391089 DOI: 10.1074/jbc.M113.511220

Date:

16-Aug-13 Release date: 08-Jan-14

Protein chains

P04275  (VWF_HUMAN) -  von Willebrand factor from Homo sapiens

Seq: 2813 a.a.

Struc: 201 a.a.*

* PDB and UniProt seqs differ at 3 residue positions (black crosses)

DOI no: 10.1074/jbc.M113.511220

J Biol Chem 289:5565-5579 (2014)

PubMed id: 24391089

Structural basis of regulation of von Willebrand factor binding to glycoprotein Ib.

M.A.Blenner, X.Dong, T.A.Springer.

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.