The C-terminal domains of TACE weaken the inhibitory action of N-TIMP-3.
Tumor necrosis factor-alpha converting enzyme (TACE) is an ADAM (a disintegrin and metalloproteinases) that comprises an active catalytic domain and several C-terminal domains. We compare the binding affinity and association rate constants of the N-terminal domain form of wild-type tissue inhibitor of metalloproteinase (TIMP-3; N-TIMP-3) and its mutants against full-length recombinant TACE and the truncated form of its catalytic domain. We show that the C-terminal domains of TACE substantially weaken the inhibitory action of N-TIMP-3. Further probing with hydroxamate inhibitors indicates that both forms of TACE have similar active site configurations. Our findings highlight the potential role of the C-terminal domains of ADAM proteinases in influencing TIMP interactions.
Lee
Meng-Huee
MH
School of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, UK. meng.lee@uea.ac.uk
Verma
Vandana
V
Maskos
Klaus
K
Becherer
J David
JD
Knäuper
Vera
V
Dodds
Philippa
P
Amour
Augustin
A
Murphy
Gillian
G
D051722:ADAM Proteins; D000072198:ADAM17 Protein; D019943:Amino Acid Substitution; D020134:Catalytic Domain / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006877:Hydroxamic Acids / Q000494:pharmacology; D007700:Kinetics; D007930:Leucine / Q000235:genetics; D008666:Metalloendopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D011994:Recombinant Proteins / Q000187:drug effects / Q000378:metabolism; D012694:Serine / Q000235:genetics; D013912:Threonine / Q000235:genetics; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D004791:Enzyme Inhibitors; D006877:Hydroxamic Acids; D011994:Recombinant Proteins; D019717:Tissue Inhibitor of Metalloproteinase-3; D013912:Threonine; D012694:Serine; D051722:ADAM Proteins; D008666:Metalloendopeptidases; D000072198:ADAM17 Protein; D007930:Leucine