Crystal Structure of the Catalytic Domain of MMP-16/MT3-MMP: Characterization of MT-MMP Specific Features

Abstract

Membrane-type matrix metalloproteinases (MT-MMPs) have attracted strong attention, because four of them can activate a key player in the tumor scenario, proMMP-2/progelatinase A. In addition to this indirect effect on the cellular environment, these MT-MMPs degrade extracellular matrix proteins, and their overproduction is associated with tumor growth. We have solved the structure of the catalytic domain (cd) of MT3-MMP/MMP-16 in complex with the hydroxamic acid inhibitor batimastat. CdMT3-MMP exhibits a classical MMP-fold with similarity to MT1-MMP Nevertheless, it also shows unique properties such as a modified MT-specific loop and a closed S1' specificity pocket, which might help to design specific inhibitors. Some MT-MMP-specific features, derived from the crystal structures of MT-1-MMP determined previously and MT3-MMP, and revealed in recent mutagenesis experiments, explain the impaired interaction of the MT-MMPs with TIMP-1. Docking experiments with proMMP-2 show some exposed loops including the MT-loop of cdMT3-MMP involved in the interaction with the proMMP-2 pro-domain in the activation encounter complex. This model might help to understand the experimentally proven importance of the MT-loop for the activation of proMMP-2.

GPI-anchored TIMP-1 treatment renders renal cell carcinoma sensitive to FAS-meditated killing

“…TIMP-1 binds most active forms of MMPs, the exceptions being MMP-14 and MMP-16 (Brew et al, 2000;Lang et al, 2004). Following incubation of RCC-53 with 700 ng/ml of TIMP-1-GPI protein for 24 h, FACS analyses using MMP-1, MMP-2, MMP-3, specific antibodies showed, with the exception of MMP-14, an increase in mean channel fluorescence intensity (MFI) for each of the MMPs.…”

“…While not much is known about TIMP-1 association with MMP-15, binding of TIMP-1-GPI protein to MMP-16 would not be predicted to occur based on the rather poor avidity of native TIMP-1 for this protein (Lang et al, 2004). However, recent structural studies have identified the amino acids that apparently explain this low association reaction/affinity (Lang et al, 2004).…”

“…While not much is known about TIMP-1 association with MMP-15, binding of TIMP-1-GPI protein to MMP-16 would not be predicted to occur based on the rather poor avidity of native TIMP-1 for this protein (Lang et al, 2004). However, recent structural studies have identified the amino acids that apparently explain this low association reaction/affinity (Lang et al, 2004). Mutational analysis of the TIMP-1 loops critical for MMP-16 binding show that small, apparently insignificant changes in TIMP-1 can dramatically shift its inhibitory/binding characteristics (Lang et al, 2004).…”

“…However, recent structural studies have identified the amino acids that apparently explain this low association reaction/affinity (Lang et al, 2004). Mutational analysis of the TIMP-1 loops critical for MMP-16 binding show that small, apparently insignificant changes in TIMP-1 can dramatically shift its inhibitory/binding characteristics (Lang et al, 2004). In this instance, the altered stoichiometry of TIMP-1 on the cell surface appears to have been sufficient to shift its binding to MMP-16.…”

Valine-based biphenylsulphonamide matrix metalloproteinase inhibitors as tumor imaging agents

“…The recombinant catalytic domain of MT1-MMP(cMTl), MT3-MMP(cMT3) and pro-MMP-2 were produced in Liège (Lang et al, 2004). Pro-MMP-9 was purchased from R&D Systems.…”