Engineering of selective TIMPs.
Differences in proteinase susceptibility between free TIMP-1 and the TIMP-1-MMP-3 complex and mutagenesis studies suggested that the residues around the disulfide bond between Cys1 and Cys70 in TIMP-1 may interact with MMPs. The crystal structure of the complex between TIMP-1 and the catalytic domain of MMP-3 has revealed that the alpha-amino group of Cys1 bidentately chelates the catalytic zinc of MMP-3 and the Thr2 side chain occupies the S1' pocket. Generation of the N-terminal domain of TIMP-1 (N-TIMP-1) variants with 15 different amino acid substitutions for Thr2 has indicated that the nature of the side chain of residue 2 has a major effect on the affinity of N-TIMP-1 for three different MMPs (MMPs-1, -2 and -3). The results also demonstrate that the mode of binding of N-TIMP-1 residue 2 differs from the binding of the P1' residue of a peptide substrate.
Nagase
H
H
Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City 66160, USA. hnagase@kumc.edu
Meng
Q
Q
Malinovskii
V
V
Huang
W
W
Chung
L
L
Bode
W
W
Maskos
K
K
Brew
K
K
D000595:Amino Acid Sequence; D000818:Animals; D006801:Humans; D007700:Kinetics; D008666:Metalloendopeptidases / Q000737:chemistry / Q000378:metabolism*; D016296:Mutagenesis; D015202:Protein Engineering; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000737:chemistry; D019714:Tissue Inhibitor of Metalloproteinases / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology
D019715:Tissue Inhibitor of Metalloproteinase-1; D019714:Tissue Inhibitor of Metalloproteinases; D008666:Metalloendopeptidases