Global Orientation of Bound MMP-3 and N-TIMP-1 in Solution via Residual Dipolar
Couplings

Matrix MetalloProteinase-3 in complex with N-TIMP-1

MMP-3 - enzyme involved in wound healing, arthritis and some forms of cancer

N-TIMP-1 inhibitor that binds to Matrix Metalloproteinases

Residue Glutamate-202 was not mutated to Glutamine in the complex studied here.

Information obtained from PDB's Chemical Component Dictionary
at http://wwpdb-remediation.rutgers.edu/downloads.html
Downloaded on Wed Jul 13 17:18:35 2011

Information obtained from PDB's Chemical Component Dictionary
at http://wwpdb-remediation.rutgers.edu/downloads.html
Downloaded on Mon Jul 25 12:26:23 2011

Most of the backbone assignments and nearly 80% of the side chain assignments
of MMP-3 when in complex with N-TIMP-1.

Global orientation of bound MMP-3 and N-TIMP-1 in solution via residual dipolar couplings.

Surfaces of the 173 residue catalytic domain of human matrix metalloproteinase 3
(MMP-3(DeltaC)) affected by binding of the N-terminal, 126 residue inhibitory
domain of human TIMP-1 (N-TIMP-1) have been investigated using an
amide-directed, NMR-based approach. The interface was mapped by a novel method
that compares amide proton line broadening by paramagnetic Gd-EDTA in the
presence and absence of the binding partner. The results are consistent with
the X-ray model of the complex of MMP-3(DeltaC) with TIMP-1 (Gomis et al.
(1997) Nature 389, 77-81). Residues Tyr155, Asn162, Val163, Leu164, His166,
Ala167, Ala169, and Phe210 of MMP-3(DeltaC) are protected from broadening by
the Gd-EDTA probe by binding to N-TIMP-1. N-TIMP-1-induced exposure of backbone
amides of Asp238, Asn240, Gly241, and Ser244 of helix C of MMP-3(DeltaC) to
Gd-EDTA confirms that the displacement of the N-terminus of MMP-3(DeltaC)
occurs not only in the crystal but also in solution. These results validate
comparative paramagnetic surface probing as a means of mapping protein-protein
interfaces. Novel N-TIMP-1-dependent changes in hydrogen bonding near the
active site of MMP-3(DeltaC) are reported. N-TIMP-1 binding causes the amide of
Tyr223 of MMP-3(DeltaC) bound by N-TIMP-1 to exchange with water rapidly,
implying a lack of the hydrogen bond observed in the crystal structure. The
backbone amide proton of Asn162 becomes protected from rapid exchange upon
forming a complex with N-TIMP-1 and could form a hydrogen bond to N-TIMP-1.
N-TIMP-1 binding dramatically increases the rate of amide hydrogen exchange of
Asp177 of the fifth beta strand of MMP-3(DeltaC), disrupting its otherwise
stable hydrogen bond.

Arumugam S, Hemme CL, Yoshida N, Suzuki K, Nagase H, Berjanskii M, Wu B, Van Doren SR.
TIMP-1 contact sites and perturbations of stromelysin 1 mapped by NMR and a
paramagnetic surface probe.
Biochemistry. 1998 Jul 7;37(27):9650-7.

TIMP-1 contact sites and perturbations of stromelysin 1 mapped by NMR and a paramagnetic surface probe.

Matrix MetalloProteinases-3 or stromelysin 1

N-terminal domain of Tissue Inhibitor of MatrixmetalloProteinases-1

Matrix MetalloProteinase-3 catalytic domain

The Mmp-Inhibitory, N-Terminal Domain Of Human Tissue Inhibitor Of Metalloproteinases-1 (N-Timp-1), Solution Nmr, 29 Structures

Orientation In Solution Of Mmp-3 Catalytic Domain And N- Timp-1 From Residual Dipolar Couplings

Crystal Structure Of The Catalytic Domain Of Mmp-1 In Complex With The Inhibitory Domain Of Timp-1

Complex Membrane Type-1 Matrix Metalloproteinase (Mt1-Mmp) With Tissue Inhibitor Of Metalloproteinase-1 (Timp-1)

Complex Of Matrix Metalloproteinase-10 Catalytic Domain (Mmp-10cd) With Tissue Inhibitor Of Metalloproteinases-1 (Timp-1)

tissue inhibitor of metalloproteinases [Homo sapiens]

prefibroblast collagenase inhibitor [Homo sapiens]

metalloprotease-1 tissue inhibitor [Papio cynocephalus]

metalloproteinase inhibitor [Homo sapiens]

TIMP metallopeptidase inhibitor 1 [Homo sapiens]

matrix metalloproteinase-1 tissue inhibitor - baboon

metalloproteinase inhibitor 1 precursor [Macaca mulatta]

metalloproteinase inhibitor 1 precursor [Pongo abelii]

metalloproteinase inhibitor 1 precursor [Homo sapiens]

PREDICTED: metalloproteinase inhibitor 1 [Nomascus leucogenys]

PREDICTED: metalloproteinase inhibitor 1 [Papio anubis]

RecName: Full=Metalloproteinase inhibitor 1; AltName: Full=Erythroid-potentiating activity; Short=EPA; AltName: Full=Fibroblast

RecName: Full=Metalloproteinase inhibitor 1; AltName: Full=Tissue inhibitor of metalloproteinases 1; Short=TIMP-1; Flags: Precu

Metalloproteinase inhibitor 1 precursor (TIMP-1)

Metalloproteinase inhibitor 1 precursor (Tissue inhibitor of metalloproteinases) (TIMP-1) (Erythroid-potentiating activity) (EPA) (Fibroblast collagenase inhibitor) (Collagenase inhibitor)

tissue inhibitor of matrix metalloproteinase-1 [Macaca mulatta]

tissue inhibitor of metalloproteinase 1 precursor [Homo sapiens]

N-terminal domain of Tissue Inhibitor of Metalloproteinases-1

N-terminal inhibitory domain of human tissue inhibitor of metalloproteinases-1

N-terminus of Tissue Inhibitor of Metalloproteinases-1/Matrix MetalloProteinases-3(catalytic domain) complex

8 MM Nalorac probe was used for most of the experiments except the
[u-2H]MMP-3/N-TIMP-1 complex where a 5 mm probe was used.

RIGID BODY MINIMIZATION FOLLOWED BY RESTRAINED SIMULATED ANNEALING

backbone and sidechain resonance assignments for N-TIMP-1