Skip to content
Licensed Unlicensed Requires Authentication Published by De Gruyter March 4, 2019

Metalloprotease inhibitor profiles of human ADAM8 in vitro and in cell-based assays

Uwe Schlomann, Kristina Dorzweiler, Elisa Nuti, Tiziano Tuccinardi, Armando Rossello and Jörg W. Bartsch ORCID logo
From the journal Biological Chemistry


ADAM8 as a membrane-anchored metalloproteinase-disintegrin is upregulated under pathological conditions such as inflammation and cancer. As active sheddase, ADAM8 can cleave several membrane proteins, among them the low-affinity receptor FcεRII CD23. Hydroxamate-based inhibitors are routinely used to define relevant proteinases involved in ectodomain shedding of membrane proteins. However, for ADAM proteinases, common hydroxamates have variable profiles in their inhibition properties, commonly known for ADAM proteinases 9, 10 and 17. Here, we determined the inhibitor profile of human ADAM8 for eight ADAM/MMP inhibitors by in vitro assays using recombinant ADAM8 as well as the in vivo inhibition in cell-based assays using HEK293 cells to monitor the release of soluble CD23 by ADAM8. ADAM8 activity is inhibited by BB94 (Batimastat), GW280264, FC387 and FC143 (two ADAM17 inhibitors), made weaker by GM6001, TAPI2 and BB2516 (Marimastat), while no inhibition was observed for GI254023, an ADAM10 specific inhibitor. Modeling of inhibitor FC143 bound to the catalytic sites of ADAM8 and ADAM17 reveals similar geometries in the pharmacophoric regions of both proteinases, which is different in ADAM10 due to replacement in the S1 position of T300 (ADAM8) and T347 (ADAM17) by V327 (ADAM10). We conclude that ADAM8 inhibitors require maximum selectivity over ADAM17 to achieve specific ADAM8 inhibition.

Funding source: Deutsche Forschungsgemeinschaft

Award Identifier / Grant number: BA1606/3-1

Funding statement: This work was supported by the Deutsche Forschungsgemeinschaft (Funder Id: 10.13039/501100001659, BA1606/3-1 to J.W.B. and U.S.) and by the University of Pisa (PRA_2018_20). We thank Sarah Koch (Marburg) for help with enzyme assays, Luciana Marinelli and Valeria La Pietra (Naples) for providing us with the docking model ADAM17/FC143, Muriel Bartsch for help with IC50 calculations, and Vincent Dive (Gif-sur-Yvette, France) for helpful discussions on the ADAM8 structure.


Almahdy, A., Koller, G., Sauro, S., Bartsch, J.W., Sherriff, M., Watson, T.F., and Banerjee, A. (2012). Effects of MMP inhibitors incorporated within dental adhesives. J. Dent. Res. 91, 605–611.10.1177/0022034512446339Search in Google Scholar

Bartsch, J.W., Wildeboer, D., Koller, G., Naus, S., Rittger, A., Moss, M.L., Minai, Y., and Jockusch, H. (2010). Tumor necrosis factor-α (TNF-α) regulates shedding of TNF-α receptor 1 by the metalloprotease-disintegrin ADAM8: evidence for a protease-regulated feedback loop in neuroprotection. J. Neurosci. 30, 12210–12218.10.1523/JNEUROSCI.1520-10.2010Search in Google Scholar

Berman, H.M., Westbrook, J., Feng, Z., Gilliland, G., Bhat, T.N., Weissig, H., Shindyalov, I.N., and Bourne, P.E. (2000). The Protein Data Bank. Nucleic Acids Res. 28, 235–242.10.1093/nar/28.1.235Search in Google Scholar

Camodeca, C., Nuti, E., Tepshi, L., Boero, S., Tuccinardi, T., Stura, E.A., Poggi, A., Zocchi, M.R., and Rossello, A. (2016). Discovery of a new selective inhibitor of A Disintegrin And Metalloprotease 10 (ADAM-10) able to reduce the shedding of NKG2D ligands in Hodgkin’s lymphoma cell models. Eur. J. Med. Chem. 111, 193–201.10.1016/j.ejmech.2016.01.053Search in Google Scholar

Conrad, C., Götte, M., Schlomann, U., Roessler, M., Pagenstecher, A., Anderson, P., Preston, J., Pruessmeyer, J., Ludwig, A., Li, R., et al. (2018). ADAM8 expression in breast cancer derived brain metastases: functional implications on MMP-9 expression and transendothelial migration in breast cancer cells. Int. J. Cancer 142, 779–791.10.1002/ijc.31090Search in Google Scholar

Dong, F., Eibach, M., Bartsch, J.W., Dolga, A.M., Schlomann, U., Conrad, C., Schieber, S., Schilling, O., Biniossek, M.L., Culmsee, C., et al. (2015). The metalloprotease-disintegrin ADAM8 contributes to temozolomide chemoresistance and enhanced invasiveness of human glioblastoma cells. Neuro. Oncol. 17, 1474–1485.10.1093/neuonc/nov042Search in Google Scholar

Fellmann, M., Buschor, P., Röthlisberger, S., Zellweger, F., and Vogel, M. (2015). High affinity targeting of CD23 inhibits IgE synthesis in human B cells. Immun. Inflamm. Dis. 3, 339–349.10.1002/iid3.72Search in Google Scholar

Fourie, A.M., Coles, F., Moreno, V., and Karlsson, L. (2003). Catalytic activity of ADAM8, ADAM15, and MDC-L (ADAM28) on synthetic peptide substrates and in ectodomain cleavage of CD23. J. Biol. Chem. 278, 30469–30477.10.1074/jbc.M213157200Search in Google Scholar

Goddard, T.D., Huang, C.C., Meng, E.C., Pettersen, E.F., Couch, G.S., Morris, J.H., and Ferrin, T.E. (2018). UCSF ChimeraX: Meeting modern challenges in visualization and analysis. Protein. Sci. 27, 14–25.10.1002/pro.3235Search in Google Scholar

Gómez-Gaviro, M., Domínguez-Luis, M., Canchado, J., Calafat, J., Janssen, H., Lara-Pezzi, E., Fourie, A., Tugores, A., Valenzuela-Fernández, A., Mollinedo, F., et al. (2007). Expression and regulation of the metalloproteinase ADAM-8 during human neutrophil pathophysiological activation and its catalytic activity on L-selectin shedding. J. Immunol. 178, 8053–8063.10.4049/jimmunol.178.12.8053Search in Google Scholar

Ishikawa, N., Daigo, Y., Yasui, W., Inai, K., Nishimura, H., Tsuchiya, E., Kohno, N., and Nakamura, Y. (2004). ADAM8 as a novel serological and histochemical marker for lung cancer. Clin. Cancer Res. 10, 8363–8370.10.1158/1078-0432.CCR-04-1436Search in Google Scholar

Koller, G., Schlomann, U., Golfi, P., Ferdous, T., Naus, S., and Bartsch, J.W. (2009). ADAM8/MS2/CD156, an emerging drug target in the treatment of inflammatory and invasive pathologies. Curr. Pharm. Des. 15, 2272–2281.10.2174/138161209788682361Search in Google Scholar

Li, C., Cantor, W.J., Nili, N., Robinson, R., Fenkell, L., Tran, Y.L., Whittingham, H.A., Tsui, W., Cheema, A.N., Sparkes, J.D., et al. (2002). Arterial repair after stenting and the effects of GM6001, a matrix metalloproteinase inhibitor. J. Am. Coll. Cardiol. 39, 1852–1858.10.1016/S0735-1097(02)01873-9Search in Google Scholar

Ludwig, A., Hundhausen, C., Lambert, M.H., Broadway, N., Andrews, R.C., Bickett, D.M., Leesnitzer, M.A., and Becherer, J.D. (2005). Metalloproteinase inhibitors for the disintegrin-like metalloproteinases ADAM10 and ADAM17 that differentially block constitutive and phorbol ester-inducible shedding of cell surface molecules. Comb. Chem. High Throughput Screen 8, 161–171.10.2174/1386207053258488Search in Google Scholar

Maretzky, T., Swendeman, S., Mogollon, E., Weskamp, G., Sahin, U., Reiss, K., and Blobel, C.P. (2017). Characterization of the catalytic properties of the membrane-anchored metalloproteinase ADAM9 in cell-based assays. Biochem. J. 474, 1467–1479.10.1042/BCJ20170075Search in Google Scholar

Miyauchi, M., Koya, J., Arai, S., Yamazaki, S., Honda, A., Kataoka, K., Yoshimi, A., Taoka, K., Kumano, K., and Kurokawa, M. (2018). ADAM8 is an antigen of tyrosine kinase inhibitor-resistant chronic myeloid leukemia cells identified by patient-derived induced pluripotent stem cells. Stem Cell Rep. 10, 1115–1130.10.1016/j.stemcr.2018.01.015Search in Google Scholar

Nuti, E., Casalini, F., Avramova, S.I., Santamaria, S., Fabbi, M., Ferrini, S., Marinelli, L., La Pietra, V., Limongelli, V., Novellino, E., et al. (2010). Potent arylsulfonamide inhibitors of tumor necrosis factor-alpha converting enzyme able to reduce activated leukocyte cell adhesion molecule shedding in cancer cell models. J. Med. Chem. 53, 2622–2635.10.1021/jm901868zSearch in Google Scholar

Nuti, E., Casalini, F., Santamaria, S., Fabbi, M., Carbotti, G., Ferrini, S., Marinelli, L., La Pietra, V., Novellino, E., Camodeca, C., et al. (2013). Selective arylsulfonamide inhibitors of ADAM-17: hit optimization and activity in ovarian cancer cell models. J. Med. Chem. 56, 8089–8103.10.1021/jm4011753Search in Google Scholar

Pettersen, E.F., Goddard, T.D., Huang, C.C., Couch, G.S., Greenblatt, D.M., Meng, E.C., and Ferrin, T.E. (2004). UCSF Chimera – a visualization system for exploratory research and analysis. J. Comput. Chem. 25, 1605–1612.10.1002/jcc.20084Search in Google Scholar

Reiss, K. and Saftig, P. (2009). The “a disintegrin and metalloprotease” (ADAM) family of sheddases: physiological and cellular functions. Semin Cell Dev. Biol. 20, 126–137.10.1016/j.semcdb.2008.11.002Search in Google Scholar

Romagnoli, M., Mineva, N.D., Polmear, M., Conrad, C., Srinivasan, S., Loussouarn, D., Barillé-Nion, S., Georgakoudi, I., Dagg, Á., McDermott, E.W., et al. (2014). ADAM8 expression in invasive breast cancer promotes tumor dissemination and metastasis. EMBO Mol. Med. 6, 278–294.10.1002/emmm.201303373Search in Google Scholar

Schlomann, U., Koller, G., Conrad, C., Ferdous, T., Golfi, P., Garcia, A.M., Höfling, S., Parsons, M., Costa, P., Soper, R., et al. (2015). ADAM8 as a drug target in pancreatic cancer. Nat. Commun. 6, 6175.10.1038/ncomms7175Search in Google Scholar

Schwarz, N., Pruessmeyer, J., Hess, F.M., Dreymueller, D., Pantaler, E., Koelsch, A., Windoffer, R., Voss, M., Sarabi, A., Weber, C., et al. (2010). Requirements for leukocyte transmigration via the transmembrane chemokine CX3CL1. Cell Mol. Life Sci. 67, 4233–4248.10.1007/s00018-010-0433-4Search in Google Scholar

Seals, D.F. and Courtneidge, S.A. (2003). The ADAMs family of metalloproteases: multidomain proteins with multiple functions. Genes Dev. 17, 7–30.10.1101/gad.1039703Search in Google Scholar

Seegar, T.C.M., Killingsworth, L.B., Saha, N., Meyer, P.A., Patra, D., Zimmerman, B., Janes, P.W., Rubinstein, E., Nikolov, D.B., Skiniotis, G., et al. (2017). Structural basis for regulated proteolysis by the α-secretase ADAM10. Cell 171, 1638–1648.e7.10.1016/j.cell.2017.11.014Search in Google Scholar

Ulasov, I., Thaci, B., Sarvaiya, P., Yi, R., Guo, D., Auffinger, B., Pytel, P., Zhang, L., Kim, C.K., Borovjagin, A., et al. (2013). Inhibition of MMP14 potentiates the therapeutic effect of temozolomide and radiation in gliomas. Cancer Med. 2, 457–467.10.1002/cam4.104Search in Google Scholar

Valkovskaya, N., Kayed, H., Felix, K., Hartmann, D., Giese, N.A., Osinsky, S.P., Friess, H., and Kleeff, J. (2007). ADAM8 expression is associated with increased invasiveness and reduced patient survival in pancreatic cancer. J. Cell Mol. Med. 11, 1162–1174.10.1111/j.1582-4934.2007.00082.xSearch in Google Scholar

Weskamp, G., Ford, J.W., Sturgill, J., Martin, S., Docherty, A.J., Swendeman, S., Broadway, N., Hartmann, D., Saftig, P., Umland, S., et al. (2006). ADAM10 is a principal ‘sheddase’ of the low-affinity immunoglobulin E receptor CD23. Nat. Immunol. 7, 1293–1298.10.1038/ni1399Search in Google Scholar

Zhang, W., Wan, M., Ma, L., Liu, X., and He, J. (2013). Protective effects of ADAM8 against cisplatin-mediated apoptosis in non-small-cell lung cancer. Cell Biol. Int. 37, 47–53.10.1002/cbin.10011Search in Google Scholar

Supplementary Material

The online version of this article offers supplementary material (

Received: 2018-10-10
Accepted: 2018-12-19
Published Online: 2019-03-04
Published in Print: 2019-06-26

©2019 Walter de Gruyter GmbH, Berlin/Boston

Scroll Up Arrow