Do the isolated fibrinogen alphaC-domains form ordered oligomers?

Biophys Chem. 2004 Dec 20;112(2-3):257-66. doi: 10.1016/j.bpc.2004.07.028.

Abstract

Previous electron microscopy (EM) studies revealed that the proteolytically prepared, truncated, bovine fibrinogen alphaC-domain (Aalpha223-539 fragment) upon transfer from acidic to neutral pH formed ordered oligomers which could mimic alpha polymers of cross-linked fibrin. In this study, we demonstrated that although its recombinant analog, bAalpha224-538, as well as the full-length version of the alphaC-domain (bAalpha224-568), upon similar treatment also formed oligomers with ordered structure, both were monomeric when kept in neutral pH buffer. To search further for conditions for their oligomerization, we treated bAalpha224-568 with factor XIIIa, purified the cross-linked soluble fraction, and confirmed that it consisted of oligomers. Similar cross-linked oligomers were obtained with the recombinant human alphaC-domain (residues Aalpha221-610). In a cell adhesion assay, the adhesion of human umbilical vein endothelial cells (HUVEC) to the alphaC-domains substantially increased upon oligomerization. These results demonstrate that the recombinant alphaC-domains can form stable oligomers which may mimic properties of the alphaC-domains in cross-linked fibrin.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cattle
  • Cell Adhesion
  • Dimerization
  • Endothelium, Vascular / cytology
  • Factor XIIIa / chemistry
  • Fibrinogen / chemistry*
  • Fibrinogen / metabolism
  • Fibrinogen / ultrastructure
  • Humans
  • Peptide Fragments / chemistry*
  • Peptide Fragments / metabolism
  • Peptide Fragments / ultrastructure
  • Protein Binding
  • Protein Structure, Tertiary
  • Umbilical Veins / cytology

Substances

  • Peptide Fragments
  • fibrinogen alphaC
  • Fibrinogen
  • Factor XIIIa