Examination of Matrix Metalloproteinase-1 in Solution

Abstract

Background: Matrix metalloproteinase-1 (MMP-1) collagenolysis relies on interdomain flexibility. Results: In all high maximum occurrence conformations, the MMP-1 hemopexin-like domain residues reported responsible for binding to the collagen triple-helix are solvent exposed. Conclusion: MMP-1 in solution is poised to interact with collagen and proceed along the steps of collagenolysis. Significance: The maximum occurrence approach can evaluate the predominant domain conformations for numerous multidomain enzymes.

Transient Collagen Triple Helix Binding to a Key Metalloproteinase in Invasion and Development

“…The hypothesis that the HPX domain folds back upon the collagen triple-helix at the active site (Bode, 1995) would then offer the needed proximity of both domains to the scissile bond region (Figure 6B). The ease of rotating the orientations between the catalytic and HPX domains is suggested by the tremendous range of fluctuating orientations between the corresponding domains of MMP-1 (Cerofolini et al, 2013), which was proposed to generalize to all MMPs due to the greater length of their flexible linkers (Bertini et al, 2008). We used atomic force microscopy (AFM) of the ectodomain to investigate this hypothesis of loose tethering of the catalytic to HPX domain of MT1-MMP, as well as the range of their relative positions.…”

How to tackle protein structural data from solution and solid state: An integrated approach

“…In these highest MaxOcc conformations the residues of the hemopexin domain responsible for collagen binding are well exposed to the solution; after collagen binding by the hemopexin domain, the catalytic domain is already preferentially oriented in such a way as to easily access the collagen as well, and a rotation by around 50°is sufficient to recover the previously proposed conformation responsible for the first step of collagenolysis [184]. Thus the MaxOcc analysis suggests that already in its free form in solution the full-length MMP-1 shows a preference for conformations which are poised for interaction with collagen and thus for catalytic activity [38]. Notably, the highest MaxOcc conformations differ largely from the closed MMP-1 structures obtained by X-ray crystallography, with a MaxOcc of about 20%.…”

Biophysical Studies of Matrix Metalloproteinase/Triple-Helix Complexes

“…2) (Arnold et al, 2011; Bertini et al, 2009). Paramagnetic NMR spectroscopic and SAXS data were subsequently used to calculate the maximum occurrence (MO) of conformations of MMP-1 in solution (Cerofolini et al, 2013). …”

“…The highest MO conformations sampled by MMP-1 when free in solution appeared to be much more poised for interaction with collagen than the compact X-ray crystallographic structures (Cerofolini et al, 2013). In the conformations belonging to this cluster (i) the collagen-binding residues of the HPX domain were solvent exposed and (ii) the CAT domain was already correctly positioned for its subsequent interaction with the collagen (Fig.…”

“…The blue (dark gray in the print version) and red (gray in the print version) arrows indicate the directions of the helices hA and hC, respectively. Reproduced from Cerofolini et al (2013) by the permission of the American Society for Biochemistry and Molecular Biology. …”