The C-terminal domains of ADAMTS-4 and ADAMTS-5 promote association with N-TIMP-3

By L. Troeberg, K. Fushimi, Simone D Scilabra, Hiroyuki Nakamura, V. Dive, I. Thøgersen, J. Enghild, Hideaki Nagase. 2009

65 citations recorded by the source corpus.

We investigated whether the affinity of tissue inhibitor of metalloproteinases (TIMP)-3 for adamalysins with thrombospondin motifs (ADAMTS)-4 and ADAMTS-5 is affected by the non-catalytic ancillary domains of the enzymes. For this purpose, we first established a novel method of purifying recombinant FLAG-tagged TIMP-3 and its inhibitory N-terminal domain (N-TIMP-3) by treating transfected HEK293 cells with sodium chlorate to prevent heparan sulfate proteoglycan-mediated TIMP-3 internalization. TIMP-3 and N-TIMP-3 affinity for selected matrix metalloproteinases and forms of ADAMTS-4 and -5 lacking sequential C-terminal domains was determined. TIMP-3 and N-TIMP-3 displayed similar affinity for various matrix metalloproteinases as has been previously reported for E. coli-expressed N-TIMP-3. ADAMTS-4 and -5 were inhibited more strongly by N-TIMP-3 than by full-length TIMP-3. The C-terminal domains of the enzymes enhanced interaction with N-TIMP-3 and to a lesser extent with the full-length inhibitor. For example, N-TIMP-3 had 7.5-fold better Ki value for full-length ADAMTS-5 than for the catalytic and disintegrin domain alone. We propose that the C-terminal domains of the enzymes affect the structure around the active site, favouring interaction with TIMP-3.

Subfields: Cancer Research, Hematology, Oncology

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Cancer Research

1. Hideaki Nagase
2. K. Brew
3. D. Edwards
4. Gillian Murphy
5. R. Khalil

In Hematology

1. Hideaki Nagase
2. D. Edwards
3. Gillian Murphy

In Oncology

1. Hideaki Nagase
2. D. Edwards

Citations in the corpus

• The tissue inhibitors of metalloproteinases (TIMPs): An ancient family with structural and functional diversity (2010, background): “For example, the inhibition of the complete extracellular region of ADAM17 by TIMP-3 is 10-fold weaker than the inhibition of the truncated catalytic domain [49], whereas the presence of the C-terminal domains of ADAMTS-4 and -5 enhances their binding to TIMP-3 [174].”
• The ADAMTS (A Disintegrin and Metalloproteinase with Thrombospondin motifs) family (2015, background): “As LRP-1 also binds and internalizes TIMP-3, and as this interaction is blocked by heparan sulfate GAGs, which also potentiates TIMP-3 activity, there is potential for modulating the tissue activities of ADAMTS using sulfated glycan mimics [45].”