This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Nakamura, H. Articles by Seiki, M. Search for Related Content PubMed PubMed Citation Articles by Nakamura, H. Articles by Seiki, M.

Constitutive and Induced CD44 Shedding by ADAM-Like Proteases and Membrane-Type 1 Matrix Metalloproteinase

¹ Department of Cancer Cell Research, Institute of Medical Science, University of Tokyo; ² Biopharmaceutical Department, Daiichi Fine Chemical, Takaoka; and ³ Departments of Otolaryngology and ⁴ Pathology, School of Medicine, Keio University, Tokyo, Japan

CD44 is a receptor for hyaluronan and mediates signaling that regulates complex cell behavior including cancer cell migration and invasion. Shedding of the extracellular portion of CD44 is the last step in the regulation of the molecule-releasing interaction between the ligand and cell. However, highly glycosylated forms of CD44 have hampered the identification of the exact cleavage sites for shedding and the responsible proteases. In this study, we found that expression of membrane-type 1 matrix metalloproteinase (MT1-MMP) increased shedding of the 65–70 kDa CD44H (standard form) fragments and generated two additional smaller fragments. We purified the shed fragments and identified the cleaved sites by mass spectrometry. Specific antibodies that recognize the newly exposed COOH terminus by cleavage were prepared and used to analyze shedding at each site. Shedding of the 65–70 kDa fragments was inhibited by tissue inhibitor of metalloproteinase 3 (TIMP-3) but not by TIMP-1 and TIMP-2, suggesting involvement of a disintegrin and metalloproteinase (ADAM)-like proteases, although shedding is affected by MT1-MMP. Conversely, shedding of the two smaller fragments was inhibited by TIMP-2 and TIMP-3 but not TIMP-1, suggesting involvement of MT1-MMP itself. Shed fragments cleaved at these sites were also detected in human tumor tissues. Increased shedding at one of the MT1-MMP-sensitive sites was observed in the tumor compared with the surrounding normal tissue. However, no significant difference was observed with shedding by ADAM-like proteases. Thus, the cleavage sites for the shedding of CD44H were identified for the first time, and the results provide a basis for exploring the unknown biologic roles of shedding at different sites.

This article has been cited by other articles:

				M. A. West, A. R. Prescott, K. M. Chan, Z. Zhou, S. Rose-John, J. Scheller, and C. Watts TLR ligand-induced podosome disassembly in dendritic cells is ADAM17 dependent J. Cell Biol., September 8, 2008; 182(5): 993 - 1005. [Abstract] [Full Text] [PDF]

				D. V. Rozanov, A. Y. Savinov, R. Williams, K. Liu, V. S. Golubkov, S. Krajewski, and A. Y. Strongin Molecular Signature of MT1-MMP: Transactivation of the Downstream Universal Gene Network in Cancer Cancer Res., June 1, 2008; 68(11): 4086 - 4096. [Abstract] [Full Text] [PDF]

				N. Ali and V. Knauper Phorbol Ester-induced Shedding of the Prostate Cancer Marker Transmembrane Protein with Epidermal Growth Factor and Two Follistatin Motifs 2 Is Mediated by the Disintegrin and Metalloproteinase-17 J. Biol. Chem., December 28, 2007; 282(52): 37378 - 37388. [Abstract] [Full Text] [PDF]

				L. Eshkar Sebban, D. Ronen, D. Levartovsky, O. Elkayam, D. Caspi, S. Aamar, H. Amital, A. Rubinow, I. Golan, D. Naor, et al. The Involvement of CD44 and Its Novel Ligand Galectin-8 in Apoptotic Regulation of Autoimmune Inflammation J. Immunol., July 15, 2007; 179(2): 1225 - 1235. [Abstract] [Full Text] [PDF]

				S. Langlois, C. Nyalendo, G. Di Tomasso, L. Labrecque, C. Roghi, G. Murphy, D. Gingras, and R. Beliveau Membrane-Type 1 Matrix Metalloproteinase Stimulates Cell Migration through Epidermal Growth Factor Receptor Transactivation Mol. Cancer Res., June 1, 2007; 5(6): 569 - 583. [Abstract] [Full Text] [PDF]

				K. Taniwaki, H. Fukamachi, K. Komori, Y. Ohtake, T. Nonaka, T. Sakamoto, T. Shiomi, Y. Okada, T. Itoh, S. Itohara, et al. Stroma-Derived Matrix Metalloproteinase (MMP)-2 Promotes Membrane Type 1-MMP-Dependent Tumor Growth in Mice Cancer Res., May 1, 2007; 67(9): 4311 - 4319. [Abstract] [Full Text] [PDF]

				A. M. Miller, M. J. Nolan, J. Choi, T. Koga, X. Shen, B. Y. J. T. Yue, and P. A. Knepper Lactate Treatment Causes NF-{kappa}B Activation and CD44 Shedding in Cultured Trabecular Meshwork Cells Invest. Ophthalmol. Vis. Sci., April 1, 2007; 48(4): 1615 - 1621. [Abstract] [Full Text] [PDF]

				H. Piccard, P. E. Van den Steen, and G. Opdenakker Hemopexin domains as multifunctional liganding modules in matrix metalloproteinases and other proteins J. Leukoc. Biol., April 1, 2007; 81(4): 870 - 892. [Abstract] [Full Text] [PDF]

				M. Takeda, S. Ogino, R. Umemoto, M. Sakakura, M. Kajiwara, K. N. Sugahara, H. Hayasaka, M. Miyasaka, H. Terasawa, and I. Shimada Ligand-induced Structural Changes of the CD44 Hyaluronan-binding Domain Revealed by NMR J. Biol. Chem., December 29, 2006; 281(52): 40089 - 40095. [Abstract] [Full Text] [PDF]

				N. Egawa, N. Koshikawa, T. Tomari, K. Nabeshima, T. Isobe, and M. Seiki Membrane Type 1 Matrix Metalloproteinase (MT1-MMP/MMP-14) Cleaves and Releases a 22-kDa Extracellular Matrix Metalloproteinase Inducer (EMMPRIN) Fragment from Tumor Cells J. Biol. Chem., December 8, 2006; 281(49): 37576 - 37585. [Abstract] [Full Text] [PDF]

				S. Misra, B. P. Toole, and S. Ghatak Hyaluronan Constitutively Regulates Activation of Multiple Receptor Tyrosine Kinases in Epithelial and Carcinoma Cells J. Biol. Chem., November 17, 2006; 281(46): 34936 - 34941. [Abstract] [Full Text] [PDF]

				A. Y. Savinov, D. V. Rozanov, and A. Y. Strongin Mechanistic insights into targeting T cell membrane proteinase to promote islet {beta}-cell rejuvenation in type 1 diabetes FASEB J, September 1, 2006; 20(11): 1793 - 1801. [Abstract] [Full Text] [PDF]

				K. J. Garton, P. J. Gough, and E. W. Raines Emerging roles for ectodomain shedding in the regulation of inflammatory responses J. Leukoc. Biol., June 1, 2006; 79(6): 1105 - 1116. [Abstract] [Full Text] [PDF]

				L. Pelletier, P. Guillaumot, B. Freche, C. Luquain, D. Christiansen, S. Brugiere, J. Garin, and S. N. Manie {gamma}-Secretase-Dependent Proteolysis of CD44 Promotes Neoplastic Transformation of Rat Fibroblastic Cells. Cancer Res., April 1, 2006; 66(7): 3681 - 3687. [Abstract] [Full Text] [PDF]

				W. Cui, J. Z. Ke, Q. Zhang, H.-Z. Ke, C. Chalouni, and A. Vignery The intracellular domain of CD44 promotes the fusion of macrophages Blood, January 15, 2006; 107(2): 796 - 805. [Abstract] [Full Text] [PDF]

				A. Y. Savinov, D. V. Rozanov, V. S. Golubkov, F. S. Wong, and A. Y. Strongin Inhibition of Membrane Type-1 Matrix Metalloproteinase by Cancer Drugs Interferes with the Homing of Diabetogenic T Cells into the Pancreas J. Biol. Chem., July 29, 2005; 280(30): 27755 - 27758. [Abstract] [Full Text] [PDF]

				E. J. Franzmann, E. P. Reategui, K. L. Carraway, K. L. Hamilton, D. T. Weed, and W. J. Goodwin Salivary Soluble CD44: A Potential Molecular Marker for Head and Neck Cancer Cancer Epidemiol. Biomarkers Prev., March 1, 2005; 14(3): 735 - 739. [Abstract] [Full Text] [PDF]

				D. V. Rozanov, S. Sikora, A. Godzik, T. I. Postnova, V. Golubkov, A. Savinov, S. Tomlinson, and A. Y. Strongin Non-proteolytic, Receptor/Ligand Interactions Associate Cellular Membrane Type-1 Matrix Metalloproteinase with the Complement Component C1q J. Biol. Chem., November 26, 2004; 279(48): 50321 - 50328. [Abstract] [Full Text] [PDF]