Heparin-binding regions of several proteins, such as
neural cell adhesion molecule,
fibronectin,
laminin,
midkine, and anti-
thrombin III have been shown to promote
neurite extension on two-dimensional surfaces. The effect of
heparin-binding peptides on
neurite extension through three-dimensional matrices was investigated by culturing embryonic chick dorsal root ganglia (DRG) within
fibrin gels containing chemically attached
heparin-
binding peptide (HBP). The length of neurites within
fibrin gels containing cross-linked HBP was increased by more than 70% over extension through
fibrin gels containing no
peptide. The HBP sequence of
antithrombin III was incorporated into the fibrin gel as the C-terminal domain of a bidomian, chimeric
peptide; the N-terminal second domain of this
peptide contained the ∀2-
plasmin inhibitor substrate for
Factor XIIIa.
Factor XIIIa, a transglutaminase, was used to chemically attach the HBP-containing chimeric peptide to the fibrin gels during
polymerization. The amount of HBP cross-linked into the fibrin gels was determined, after degradation by
plasmin using
gel permeation chromatography, to be approximately 8 moles of peptide per mole
fibrinogen.
A peptide (HBP), where the cross-linking
glutamine was replaced with
glycine, showed no increase in extension in comparison with fibrin gels. The additional of
heparin to the gel percursors resulted in no increase in
neurite extension in comparison with fibrin gels. HBPs promote neurite extension by binding to
cell surface proteoglycans on the DRG.