Fibronectin and 130-kDa molecule complex mimics snake venom botrocetin-like structure potentially modulating association between von Willebrand factor and vascular vessel wall.
We established seven hybridomas secreting monoclonal antibodies (MoAbs) against the venom from Bothrops jararaca. Six of them were demonstrated to specifically recognize botrocetin, a venom protein which binds with von Willebrand factor (vWf) and induces platelet agglutination. Two of them, BCT4-3 and BCT115-2 MoAbs, could significantly inhibit botrocetin binding with plasma vWf. BCT4-3 could react slightly with a monolayer of human endothelial cells (ECs), and BCT4-3 binding to ECs was drastically enhanced by the coexistence of human plasma in a dose-dependent manner, indicating that a biological modulator structurally resembling botrocetin is created initially on the EC surface complexed with some plasma proteins. Botrocetin-like components could be immuno-purified only by immobilized BCT4-3, but not by the other immobilized MoAbs, from umbilical vein extracts. Interestingly, the immunoisolated materials were identified to consist essentially of fibronectin (Fn) and a 130 kDa molecule, and this complex bound to vWf in the extracts. Depletion of Fn from plasma decreased BCT4-3 binding to ECs. The epitope of BCT4-3 expressed on the endothelial surface, comprising plasma Fn and the coisolated 130 kDa molecule, is proposed to be a physiological modulator structurally mimicking botrocetin, and essentially supporting vWf-binding to injured endothelium and subsequently to circulating platelets.
Katayama
M
M
Takara Shuzo Co., Ltd., Biotechnology Research Laboratories, Shiga.
Nagata
S
S
Hirai
S
S
Miura
S
S
Fujimura
Y
Y
Matusi
T
T
Kato
I
I
Titani
K
K
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001665:Binding Sites; D001798:Blood Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002478:Cells, Cultured; D003435:Crotalid Venoms / Q000032:analysis / Q000378:metabolism* / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000187:drug effects / Q000502:physiology*; D004797:Enzyme-Linked Immunosorbent Assay; D005353:Fibronectins / Q000494:pharmacology*; D006388:Hemagglutinins / Q000494:pharmacology; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D008970:Molecular Weight; D009131:Muscle, Smooth, Vascular / Q000502:physiology; D014471:Umbilical Veins; D014841:von Willebrand Factor / Q000187:drug effects / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D001798:Blood Proteins; D003435:Crotalid Venoms; D005353:Fibronectins; D006388:Hemagglutinins; D014841:von Willebrand Factor; C030961:botrocetin