Mapping the binding site for matrix metalloproteinase on the N-terminal domain of the tissue inhibitor of metalloproteinases-2 by NMR chemical shift perturbation.
Changes in the NMR chemical shift of backbone amide nuclei (1H and 15N) have been used to map the matrix metalloproteinase (MMP) binding site on the N-terminal domain of the tissue inhibitor of metalloproteinase-2 (N-TIMP-2). Amide chemical shift changes were measured on formation of a stable complex with the catalytic domain of stromelysin-1 (N-MMP-3). Residues with significantly shifted amide signals mapped specifically to a broad site covering one face of the molecule. This site (the MMP binding site) consists primarily of residues 1-11, 27-41, 68-73, 87-90, and 97-104. The site overlaps with the OB-fold binding site seen in other proteins that share the same five-stranded beta-barrel topology. Sequence conservation data and recent site-directed mutagenesis studies are discussed in relation to the MMP binding site identified in this work.
Williamson
R A
RA
Research School of Biosciences, University of Kent, Canterbury, U.K. r.a.williamson@ukc.ac.uk
Carr
M D
MD
Frenkiel
T A
TA
Feeney
J
J
Freedman
R B
RB
D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D002247:Carbon Isotopes; D017124:Conserved Sequence / Q000235:genetics; D006801:Humans; D006859:Hydrogen; D009682:Magnetic Resonance Spectroscopy; D019278:Matrix Metalloproteinase 3 / Q000737:chemistry / Q000378:metabolism; D008666:Metalloendopeptidases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009587:Nitrogen Isotopes; D010449:Peptide Mapping / Q000379:methods*; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D019716:Tissue Inhibitor of Metalloproteinase-2; D006859:Hydrogen; D008666:Metalloendopeptidases; D019278:Matrix Metalloproteinase 3