ADAMTS-1 Protein Anchors at the Extracellular Matrix through the Thrombospondin Type I Motifs and Its Spacing Region*
- From the ‡Department of Pharmacology, Cancer Research Institute, Kanazawa University, 13-1 Takara-machi, Kanazawa, Ishikawa 920, Japan and the §Department of Molecular Preventive Medicine, School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113 Japan
Abstract
Cellular disintegrin and metalloproteinases (ADAMs) are a family of genes with a sequence similar to those of snake venom metalloproteinases and disintegrins. The ADAMTS-1 gene encodes a new type of ADAM protein with respect to possessing the thrombospondin (TSP) type I motifs. Expression of the gene is induced in kidney and heart by in vivo administration of lipopolysaccharide, suggesting a possible role in the inflammatory reaction. In this study, we characterized the ADAMTS-1 gene product by using a transient expression system in COS-7 cells. We found that the precursor and processed forms of ADAMTS-1 were secreted from cells. Under normal growth conditions, little or none of both forms was detected in the cell culture medium, and instead the majority was found associated with the extracellular matrix (ECM). In addition, when cells were cultured in the presence of heparin, the mature form of ADAMTS-1 protein was detected in the cell culture medium, suggesting that binding of ADAMTS-1 to the ECM is mediated through sulfated glycosaminoglycans such as heparan sulfate. Analyses of deletion mutants of the ADAMTS-1 protein revealed that the spacer region as well as three TSP type I motifs in the carboxyl-terminal region of the ADAMTS-1 protein are important for a tight interaction with the ECM. These results suggest that the ADAMTS-1 is a unique ADAM family protein that anchors at the ECM.
Footnotes
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↵* The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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↵¶ To whom correspondences should be addressed. Tel.: 81-03-3812-2111 (ext. 3431); Fax: 81-03-5684-2297; E-mail:koujim{at}m.u-tokyo.ac.jp
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↵1 The abbreviations used are: ADAM(s), disintegrin and metalloproteinase(s); DMEM, Dulbecco’s modified Eagle’s medium; ECM, extracellular matrix; GST, glutathione S-transferase; HPLC, high performance liquid chromatography; MDC, metalloproteinase-like/disintegrin-like/cysteine-rich; PCR, polymerase chain reaction; TNF-α, tumor necrosis factor α; TACE, tumor necrosis factor α-converting enzyme; TSP, thrombospondin; DPM,deletion of proprotein andmetalloproteinase domains.
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↵2 K. Kuno and K. Matsushima, unpublished results.
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- Received January 15, 1998.
- Revision received March 24, 1998.
- The American Society for Biochemistry and Molecular Biology, Inc.
