To compare the substrate preferences of rat brain
neurolysin and
cancer-producing
matrix metalloproteinases (
MMPs), which have the same architecture in their catalytic domains, the cleavage activity of
neurolysin toward
MMP-specific fluorescence-quenching
peptides was quantitatively measured. The results show that
neurolysin effectively cleaved MOCAc [(7-methoxy
coumarin-4-yl) acetyl]-RPKPYANvaWMK(Dnp[2,4-dinitrophenyl])-NH(2), a specific substrate of MMP-2 and MMP-9, but hardly cleaved MOCAc-RPKPVENvaWRK(Dnp)-NH(2), a specific substrate of MMP-3, suggesting that
neurolysin has a similar substrate preference to MMP-2 and MMP-9. A structural comparison between
neurolysin and MMP-9 showed the similar key
amino acid residues for substrate recognition. The possible application of
neurolysin displayed on the yeast cell surface, as a safe
protein alternative to MMP-2 and MMP-9 which induce
cancer cell growth, invasion, and
metastasis, to analysis of properties of the
MMPs, including the screening of inhibitors and analysis of inhibition mechanism etc., are also discussed.