The C-terminal domains of ADAMTS-4 and ADAMTS-5 promote association with N-TIMP-3.
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...
| Glavni autori: | , , , , , , , |
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| Format: | Journal article |
| Jezik: | English |
| Izdano: |
2009
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| _version_ | 1826288214921445376 |
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| author | Troeberg, L Fushimi, K Scilabra, S Nakamura, H Dive, V Thøgersen, I Enghild, J Nagase, H |
| author_facet | Troeberg, L Fushimi, K Scilabra, S Nakamura, H Dive, V Thøgersen, I Enghild, J Nagase, H |
| author_sort | Troeberg, L |
| collection | OXFORD |
| description | 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 K(i) 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. |
| first_indexed | 2024-03-07T02:10:20Z |
| format | Journal article |
| id | oxford-uuid:a068a4b3-3b45-4c46-9e6a-0ae87431962b |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T02:10:20Z |
| publishDate | 2009 |
| record_format | dspace |
| spelling | oxford-uuid:a068a4b3-3b45-4c46-9e6a-0ae87431962b2022-03-27T02:05:22ZThe C-terminal domains of ADAMTS-4 and ADAMTS-5 promote association with N-TIMP-3.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:a068a4b3-3b45-4c46-9e6a-0ae87431962bEnglishSymplectic Elements at Oxford2009Troeberg, LFushimi, KScilabra, SNakamura, HDive, VThøgersen, IEnghild, JNagase, HWe 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 K(i) 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. |
| spellingShingle | Troeberg, L Fushimi, K Scilabra, S Nakamura, H Dive, V Thøgersen, I Enghild, J Nagase, H The C-terminal domains of ADAMTS-4 and ADAMTS-5 promote association with N-TIMP-3. |
| title | The C-terminal domains of ADAMTS-4 and ADAMTS-5 promote association with N-TIMP-3. |
| title_full | The C-terminal domains of ADAMTS-4 and ADAMTS-5 promote association with N-TIMP-3. |
| title_fullStr | The C-terminal domains of ADAMTS-4 and ADAMTS-5 promote association with N-TIMP-3. |
| title_full_unstemmed | The C-terminal domains of ADAMTS-4 and ADAMTS-5 promote association with N-TIMP-3. |
| title_short | The C-terminal domains of ADAMTS-4 and ADAMTS-5 promote association with N-TIMP-3. |
| title_sort | c terminal domains of adamts 4 and adamts 5 promote association with n timp 3 |
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