1OO9

Orientation in Solution of MMP-3 Catalytic Domain and N-TIMP-1 from Residual Dipolar Couplings

Summary for 1OO9

• Entry DOI: 10.2210/pdb1oo9/pdb
• Related: 1UEA
• NMR Information: BMRB: 5785
• Descriptor: Stromelysin-1, Metalloproteinase inhibitor 1 (2 entities in total)
• Functional Keywords: protein-protein complex, hydrolase
• Biological source: Homo sapiens (human); More
• Cellular location: Secreted, extracellular space, extracellular matrix (Probable): P08254; Secreted: P01033
• Total number of polymer chains: 2
• Total formula weight: 33156.34

Authors

Arumugam, S.,Van Doren, S.R. (deposition date: 2003-03-03, release date: 2003-07-29, Last modification date: 2024-11-20)

Primary citation

Arumugam, S.,Van Doren, S.R.
Global Orientation of Bound MMP-3 and N-TIMP-1 in Solution via Residual Dipolar Couplings
Biochemistry, 42:7950-7958, 2003

PubMed Abstract: Crystal structures of catalytic domains of MMP-3 and MT1-MMP bound to TIMP-1 or TIMP-2, respectively, differ in the orientation of the TIMP in the MMP active site. The orientation in solution of N-TIMP-1 in the MMP-3 active site has been investigated using residual dipolar couplings (RDCs). Fitting of the RDCs to the X-ray structures of the complexes suggests general agreement with the orientation of crystalline MMP-3(DeltaC) and TIMP-1 and a large disparity from the orientation of crystalline MT1-MMP(DeltaC) and TIMP-2. Rigid body docking of MMP-3 and N-TIMP-1 X-ray coordinates using RDCs and intermolecular NOEs provided a time-averaged orientation in solution differing from the crystal structure by a 5 degrees rotation toward the MT1-MMP(DeltaC)/TIMP-2 orientation. The slight discrepancy in orientations in solution and crystal lies within the experimental uncertainties. Intermolecular NOEs used in the docking corroborated the accuracy of mapping the interface by a paramagnetic NMR footprinting assay, a potential alternative source of contacts for docking. Some uncertainty in the N-TIMP-1 orientation in the MMP-3 active site, coupled with microsecond to millisecond fluctuations of the MMP-binding ridge of N-TIMP-1 in the complex and flexibility in MMP-3(DeltaC) S(1)' to S(3)' subsites, leaves open the possibility that N-TIMP-1 might dynamically pivot a few degrees or more in the arc toward the MT1-MMP(DeltaC)/TIMP-2 orientation. Differing TIMP orientations in MMP active sites are more likely to result from structural differences in TIMP AB hairpin loops than from crystal packing artifacts.

PubMed: 12834347
DOI: 10.1021/bi034545s

Experimental method

SOLUTION NMR