Recombinant fibrinogen Vlissingen/Frankfurt IV. The deletion of residues 319 and 320 from the gamma chain of firbinogen alters calcium binding, fibrin polymerization, cross-linking, and platelet aggregation.
We synthesized a variant, recombinant fibrinogen modeled after the heterozygous dysfibrinogen Vlissingen/Frankfurt IV, a deletion of two residues, gammaAsn-319 and gammaAsp-320, located within the high affinity calcium-binding pocket. Turbidity studies showed no evidence of fibrin polymerization, although size exclusion chromatography, transmission electron microscopy, and dynamic light scattering studies showed small aggregates. These aggregates did not resemble normal protofibrils nor did they clot. Fibrinopeptide A release was normal, whereas fibrinopeptide B release was delayed approximately 3-fold. Plasmin cleavage of this fibrinogen was not changed by the presence of calcium or Gly-Pro-Arg-Pro, indicating that both the calcium-binding site and the "a" polymerization site were non-functional. We conclude that the loss of normal polymerization was due to the lack of "A-a" interactions. Moreover, functions associated with the C-terminal end of the gamma chain, such as platelet aggregation and factor XIII cross-linking, were also disrupted, suggesting that this deletion of two residues affected the overall structure of the C-terminal domain of the gamma chain.
Hogan
K A
KA
Department of Pathology and Laboratory Medicine, University of North Carolina, Chapel Hill, North Carolina 27599-7525, USA.
Gorkun
O V
OV
Lounes
K C
KC
Coates
A I
AI
Weisel
J W
JW
Hantgan
R R
RR
Lord
S T
ST
D002118:Calcium / Q000378:metabolism*; D005337:Fibrin / Q000378:metabolism*; D015241:Fibrinogens, Abnormal / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D006579:Heterozygote; D006801:Humans; D007700:Kinetics; D008854:Microscopy, Electron; D010974:Platelet Aggregation / Q000502:physiology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D017384:Sequence Deletion
D015241:Fibrinogens, Abnormal; D011994:Recombinant Proteins; C069866:fibrinogen Vlissingen; C410729:fibrinogen Vlissingen-Frankfurt IV; D005337:Fibrin; D002118:Calcium