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PMID:16912226

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Citation

Lisman, T, Raynal, N, Groeneveld, D, Maddox, B, Peachey, AR, Huizinga, EG, de Groot, PG and Farndale, RW (2006) A single high-affinity binding site for von Willebrand factor in collagen III, identified using synthetic triple-helical peptides. Blood 108:3753-6

Abstract

The essential event in platelet adhesion to the injured blood vessel wall is the binding to subendothelial collagen of plasma von Willebrand factor (VWF), a protein that interacts transiently with platelet glycoprotein Ibalpha (GPIbalpha), slowing circulating platelets to facilitate firm adhesion through collagen receptors, including integrin alpha2beta1 and GpVI. To locate the site in collagen that binds VWF, we synthesized 57 overlapping triple-helical peptides comprising the whole triple-helical domain of collagen III. Peptide no. 23 alone bound VWF, with similar affinity to that of native collagen III. Immobilized peptide no. 23 supported platelet adhesion under static and flow conditions, processes blocked by an antibody that prevents collagen from binding the VWF A3 domain. Truncated and alanine-substituted peptides derived from no. 23 either strongly interacted with both VWF and platelets or lacked both VWF and platelet binding. Thus, we identified the sequence RGQOGVMGF (O is hydroxyproline) as the minimal VWF-binding sequence in collagen III.

Links

PubMed Online version:10.1182/blood-2006-03-011965

Keywords

Amino Acid Motifs; Antibodies/chemistry; Antibodies/metabolism; Binding Sites; Collagen Type III/chemistry; Collagen Type III/metabolism; Humans; Integrin alpha2beta1/metabolism; Peptides/chemical synthesis; Peptides/chemistry; Peptides/metabolism; Platelet Adhesiveness/physiology; Platelet Glycoprotein GPIb-IX Complex/metabolism; Platelet Membrane Glycoproteins/metabolism; Protein Binding; von Willebrand Factor/chemistry; von Willebrand Factor/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:CO3A1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P04275

F

Seeded From UniProt

complete

HUMAN:VWF

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P02461

F

Seeded From UniProt

complete

HUMAN:CO3A1

involved_in

GO:0007160: cell-matrix adhesion

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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