Selective inhibition of ADAM12 catalytic activity through engineering of tissue inhibitor of metalloproteinases (TIMP)-2 - Archive ouverte HAL Access content directly
Journal Articles Biochemical Journal Year : 2010

Selective inhibition of ADAM12 catalytic activity through engineering of tissue inhibitor of metalloproteinases (TIMP)-2

Jonas Jacobsen
  • Function : Author
Meng-Huee Lee
  • Function : Author
Hideaki Nagase
  • Function : Author
Ulla M Wewer
  • Function : Author
Gillian Murphy
  • Function : Author

Abstract

The disintegrin and metalloprotease ADAM12 has important functions in normal physiology as well as in diseases, such as cancer. Little is known about how ADAM12 confers its protumorigenic effect; however, its proteolytic capacity is likely a key component. Thus, selective inhibition of ADAM12 activity may be of great value therapeutically and as an investigative tool to elucidate its mechanisms of actions. We have previously reported the inhibitory profile of tissue inhibitors of metalloproteinases (TIMPs) against ADAM12, demonstrating in addition to TIMP-3, a unique ADAM inhibitory activity of TIMP-2. These findings strongly suggest that it is feasible to design a TIMP mutant selectively inhibiting ADAM12. With this purpose, we characterized the molecular determinants of the ADAM12–TIMP complex formation as compared with known molecular requirements for TIMP-mediated inhibition of ADAM17/TACE. Kinetic analysis using a fluorescent-peptide substrate demonstrated that the molecular interactions of N-terminal domains of TIMPs (N-TIMPs) with ADAM12 and TACE are for the most part comparable, yet revealed striking unique features of TIMP-mediated ADAM12 inhibition. Intriguingly, we found that removal of the AB-loop in N-TIMP-2, which is known to impair its interaction with TACE, resulted in increased affinity to ADAM12. Importantly, using a cell-based epidermal growth factor-shedding assay, we demonstrated for the first time an inhibitory activity of TIMPs against the transmembrane ADAM12-L, verifying the distinctive inhibitory abilities of N-TIMP-2 and engineered N-TIMP-2 mutants in a cellular environment. Together, our findings support the idea that a distinctive ADAM12 inhibitor with future therapeutic potential can be designed.

Keywords

Fichier principal
Vignette du fichier
PEER_stage2_10.1042%2FBJ20100649.pdf (622.82 Ko) Télécharger le fichier
Origin : Files produced by the author(s)
Loading...

Dates and versions

hal-00506526 , version 1 (28-07-2010)

Identifiers

Cite

Marie Kveiborg, Jonas Jacobsen, Meng-Huee Lee, Hideaki Nagase, Ulla M Wewer, et al.. Selective inhibition of ADAM12 catalytic activity through engineering of tissue inhibitor of metalloproteinases (TIMP)-2. Biochemical Journal, 2010, 430 (1), pp.79-86. ⟨10.1042/BJ20100649⟩. ⟨hal-00506526⟩

Collections

PEER
62 View
209 Download

Altmetric

Share

Gmail Facebook X LinkedIn More