Reduced platelet adhesion in flowing blood to fibrinogen by alterations in segment gamma316-322, part of the fibrin-specific region.
The interaction of platelets with fibrinogen is a key event in the maintenance of a haemostatic response. It has been shown that the 12-carboxy-terminal residues of the gamma-chain of fibrinogen mediate platelet adhesion to immobilized fibrinogen. These studies, however, did not exclude the possibility that other domains of fibrinogen are involved in interactions with platelets. To obtain more insight into the involvement of other domains of fibrinogen in platelet adhesion, we studied platelet adhesion in flowing blood to patient dysfibrinogen Vlissingen/Frankfurt IV (V/FIV), to several variant recombinant fibrinogens with abnormalities in the gamma-chain segments gamma318-320 and gamma408-411. Perfusion studies at physiological shear rates showed that platelet adhesion was absent to gammaDelta408-411, slightly reduced to the heterozygous patient dysfibrinogen V/FIV and strongly reduced to the homozygous recombinant fibrinogens: gammaDelta319-320, gamma318Asp-->Ala and gamma320Asp-->Ala. Furthermore, antibodies raised against the sequences gamma308-322 and gamma316-333 inhibited platelet adhesion under shear conditions. These experiments indicated that the overlapping segment gamma316-322 contains amino acids that could be involved in platelet adhesion to immobilized fibrinogen under flow conditions. In soluble fibrinogen, this sequence is buried inside the fibrinogen molecule and becomes exposed after polymerization. In addition, we have shown that this fibrin-specific sequence also becomes exposed when fibrinogen is immobilized on a surface.
Remijn
Jasper A
JA
Thrombosis and Haemostasis Laboratory, Department of Haematology, University Medical Centre Utrecht, PO Box 85500, 3508 GA Utrecht, The Netherlands. J.A.Remijn@lab.azu.nl
IJsseldijk
Martin J W
MJ
van Hemel
Bettien M
BM
Galanakis
Dennis K
DK
Hogan
Kelly A
KA
Lounes
Karim C
KC
Lord
Susan T
ST
Sixma
Jan J
JJ
de Groot
Philip G
PG
D000911:Antibodies, Monoclonal / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005337:Fibrin / Q000235:genetics*; D005340:Fibrinogen / Q000235:genetics* / Q000276:immunology / Q000502:physiology; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology; D010973:Platelet Adhesiveness; D011994:Recombinant Proteins
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D010446:Peptide Fragments; D011994:Recombinant Proteins; D005337:Fibrin; D005340:Fibrinogen