Structure of the catalytic domain of human fibroblast collagenase complexed with an inhibitor

Abstract

In rheumatoid and osteoarthritis, degradation of articular cartilage is mediated by the matrix metalloproteinases collagenase, stromelysin and gelatinase. The key event in this process is the cleavage of triple helical collagen by collagenase. We have determined the crystal structure of the catalytic domain of human recombinant fibroblast collagenase complexed with a synthetic inhibitor at 2.2 A resolution. The protein fold is similar to the amino termini of the zinc endopeptidases astacin thermolysin and elastase despite a lack of primary sequence homology. The conformation of the bound inhibitor provides a molecular basis for the design of inhibitors of collagenase and other matrix metalloproteinases. Such inhibitors should be useful in the treatment of a variety of diseases including arthritis and cancer.

Structure of malonic acid‐based inhibitors bound to human neutrophil collagenase. A new binding mode explains apparently anomalous data

“…Most of these mimic peptides and most likely bind analogous to the corresponding peptide substrates (Fig. l), as has been shown in several X-ray crystallographic studies Borkakoti et al, 1994;Lovejoy et al, 1994;Reinemer et al, 1994;Spurlino et al, 1994;Stams et al, 1994;Browner et al, 1995;Grams et al, 1995aGrams et al, , 1995bDhanaraj et ai., 1996). The generalized structures of most of these inhibitors have been previously described (Beckett et al, 1996;Fig.…”

The Structure of the Aeromonas proteolytica Aminopeptidase Complexed with a Hydroxamate Inhibitor

“…A similar involvement of Glul43 in the binding of HONH-isobutylmalonyl-Ala-Gly-NH, to thermolysin was established from kinetic measurements (Izquierdo-Martin and Stein, 1992). Importantly, in all the previously reported structures of metallopeptidases complexed with a hydroxamate inhibitor, the interactions of an acidic glutamic acid via its carboxyl oxygens with the hydroxylamino nitrogen and oxygen appears as a general common feature (Holmes and Matthews, 1981;Borkakoti et al, 1994;Spurlino et al, 1994;Bode et al, 1994;Stams et al, 1994). The conservation of a glutamic acid residue in the active site suggests that the hydrolytic mechanism of these metalloproteases could be closely related.…”

“…In hydroxamate complexes of thermolysin (Holmes and Matthews, 1981), human fibroblast collagenase (Borkakoti et al, 1994;Spurlino et al, 1994) and human neutrophil collagenase (Bode et al, 1994;Stams et al, 1994), the catalytic zinc ion is pentacoordinated by three protein ligands and by the hydroxyl and carbonyl oxygen atoms of the hydroxamates, which act unambiguously as bidentate ligands. Our data show similarities with hydroxamate one-zinc endopeptidase complexes.…”

Specific amino acid substitutions in human collagenase cause decreased autoproteolysis and reveal a requirement for a second zinc atom for catalytic activity

“…We have also investigated the function of the second noncatalytic zinc atom [l 11, which has been confirmed in the crystal structure of collagenase domain II [12,13]. Analysis of the structure revealed the chelating groups to be H 149, D15 1, H164 and H177.…”