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Evaluation of clinical colon carcinoma using activity-based proteomic profiling

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The evaluation of clinical tumor tissues is a valuable approach to discovering novel drug targets because of the direct relevance of human samples. We used activity-based proteomic profiling (ABPP) to study the differences in serine hydrolase activities from 12 matched pairs of clinical normal and tumor colon tissues. Unlike traditional proteomics or measures of mRNA abundance, ABPP actually quantifies enzymatic activities, a characteristic crucial for drug targeting. Several serine hydrolases were differentially active in tumor vs normal tissues, despite a lack of obvious corresponding alterations in protein expression. We identified one tumor-specific activity by mass spectrometry to be fibroblast activation protein (FAP), an integral membrane serine protease that has been reported to be present only in tumor stroma or during wound healing and absent in normal tissues. FAP activity was further found to be approximately twofold higher in stage III relative to stage II colon cancer, suggestive of a role in tumor progression. We were also able to identify other proteins, some of which had not been previously linked to cancer, which had higher activity in tumors. Our results demonstrate the applicability of ABPP for the efficient identification of multiple clinical disease targets.


  1. 1.

    American Cancer Society. Cancer facts and figures 2004. 2004. Available at: Accessed on 8/19/04.

  2. 2.

    Patricelli, M.P., Giang, D.K., Stamp, L.M., and Burbaum, J.J. (2001). Direct visualization of serine hydrolase activities in complex proteomes using fluorescent active site-directed probes. Proteomics 1:1067–1071.

  3. 3.

    Liu, Y., Patricelli, M.P., and Cravatt, B.F. (1999). Activity-based protein profiling: the serine hydrolases. Proc. Natl. Acad. Sci. USA 96:14694–14699.

  4. 4.

    Hanahan, D. and Weinberg, R.A. (2000). The hallmarks of cancer. Cell 100:57–70.

  5. 5.

    Martin, M., Pujuguet, P., and Martin, F. (1996). Role of stromal myofibroblasts infiltrating colon cancer in tumor invasion. Pathol. Res. Pract. 192:712–717.

  6. 6.

    Schurch W. (1999). The myofibroblast in neoplasia. Curr. Top. Pathol. 93:135–148.

  7. 7.

    Kuhajda, F.P., Pizer, E.S., Li, J.N., Mani, N.S., Frehywot, G.L., and Townsend, C.A. Synthesis and antitumor activity of an inhibitor of fatty acid synthase. Proc. Natl. Acad. Sci. USA 97:3450–3454.

  8. 8.

    Li, J.N., Gorospe, M., Chrest, F.J., Kumaravel, T.S., Evans, M.K., Han, W.F., et al. (2001). Pharmacological inhibition of fatty acid synthase activity produces both cytostatic and cytotoxic effects modulated by p53. Cancer Res. 61:1493–1499.

  9. 9.

    Holst, J.J. and Deacon, C.F. (1998). Inhibition of the activity of dipeptidyl-peptidase IV as a treatment for type 2 diabetes. Diabetes 47:1663–1670.

  10. 10.

    Weideman, P.E. and Trevillyan, J.M. (2003). Dipeptidyl peptidase IV inhibitors for the treatment of impaired glucose tolerance and type 2 diabetes. Curr. Opin. Investig. Drugs 4:412–420.

  11. 11.

    Toide, K., Shinoda, M., and Miyazaki, A. (1998). A novel prolyl endopeptidase inhibitor, JTP-4819—its behavioral and neurochemical properties for the treatment of Alzheimer's disease. Rev. Neurosci. 9:17–29.

  12. 12.

    Bellemère, G., Morain, P., Vaudry, H., and Jegou, S. (2003). Effect of S 17092, a novel prolyl endopeptidase inhibitor, on substance P and alpha-melanocyte-stimulating hormone breakdown in the rat brain. J. Neurochem. 84: 919–929.

  13. 13.

    Šedo, A., Křepela, E., and Kasafirek, E. (1991). Dipeptidyl peptidase IV, prolyl endopeptidase and cathepsin B activities in primary human lung tumors and lung parenchyma. J. Cancer Res. Clin. Oncol. 117:249–253.

  14. 14.

    de la Haba-Rodriguez, J., Macho, A., Calzado, M.A., Blazquez, M.V., Gomez, M.A., Munoz, E.E., et al. (2002). Soluble dipeptidyl peptidase IV (CD-26) in serum of patients with colorectal carcinoma. Neoplasma 49:307–311.

  15. 15.

    Garin-Chesa, P., Old, L.J., and Rettig, W.J. (1990). Cell surface glycoprotein of reactive stromal fibroblasts as a potential antibody target in human epithelial cancers. Proc. Natl. Acad. Sci. USA 87:7235–7239.

  16. 16.

    Park, J.E., Lenter, M.C., Zimmermann, R.N., Garin-Chesa, P., Old, L.J., and Rettig, W.J. (1999). Fibroblast activation protein, a dual specificity serine protease expressed in reactive human tumor stromal fibroblasts. J. Biol. Chem. 274:36505–36512.

  17. 17.

    Kelly, T. (1999). Evaluation of seprase activity. Clin. Exp. Metastasis 17:57–62.

  18. 18.

    Scanlan M.J., Raj, B.K., Calvo, B., Garin-Chesa, P., Sanz-Moncasi, M.P., Healey, J.H., et al. (1994). Molecular cloning of fibroblast activation protein alpha, a member of the serine protease family selectivity expressed in stromal fibroblasts of epithelial cancers. Proc. Natl. Acad. Sci. USA 91:5657–5661.

  19. 19.

    Wesley, U.V., Albino, A.P., Tiwari, S., and Houghton, A.N. (1999). A role of dipeptidyl peptidase IV in suppressing the malignant phenotype of melanocytic cells. J. Exp. Med. 190:311–322.

  20. 20.

    Rettig, W.J., Su, S.L., Fortunato, S.R., Scanlan, M.J., Raj, B.K., Garin-Chesa, P., et al. (1994). Fibroblast activation protein: purification, epitope mapping and induction by growth factors. Int. J. Cancer 58:385–392.

  21. 21.

    Okerberg, E.S., Wu, J., Zhang, B., Samii, B., Blackford K., Winn, D.T., et al. (2005). High-resolution functional proteomics by active-site peptide profiling. Proc. Natl. Acad. Sci. USA 102:4996–5001.

  22. 22.

    Sanghani, S.P., Quinney, S.K., Fredenburg, T.B., Sun, Z., Davis, W.I., Murry, D.J., et al. (2003). Carboxylesterases expressed in human colon tumor tissue and their role in CPT-11 hydrolysis. Clin. Cancer Res. 9:4983–4991.

  23. 23.

    Humerickhouse, R., Lohrbach, K., Li, L., Bosron, W.F., and Dolan, M.E. (2000). Characterization of CPT-11 hydrolysis by human liver carboxylesterase isoforms hCE-1 and hCE-2. Cancer Res. 60:1189–1192.

  24. 24.

    Xu, G., Zhang, W., Ma, M.K., and McLeod, H.L. (2002). Human carboxylesterase 2 is commonly expressed in tumor tissue and is correlated with activation of irinotecan. Clin. Cancer Res. 8:2605–2611.

  25. 25.

    Guichard, S., Terret, C., Hennebelle, I., Lochon, I., Chevreau, P., Fretigny, E., et al. (1999). CPT-11 converting carboxylesterase and topoisomerase activities in tumour and normal colon and liver tissues. Br. J. Cancer 80:364–370.

  26. 26.

    Tobin, P.J., Dodds, H.M., Clarke, S., Schnitzler, M., and Rivory, L.P. (2003). The relative contributions of carboxylesterase and beta-glucuronidase in the formation of SN-38 in human colorectal tumours. Oncol. Rep. 10: 1977–1979.

  27. 27.

    Stafforini, D.M., McIntyre, T.M., Zimmerman, G.A., and Prescott, S.M. (1997). Platelet-activating factor acetylhydrolases. J. Biol. Chem. 272:17895–17898.

  28. 28.

    Arai, H., Koizumi, H., Aoki, J., and Inoue, K. (2002). Platelet-activating factor acetylhydrolase (FAF-AH). J. Biochem. 131:635–640.

  29. 29.

    Denizot, Y., Desplat, V., Drouet, M., Bertin, F., and Melloni, B. (2001). Is there a role of platelet-activating factor in human lung cancer? Lung Cancer 33:195–202.

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Correspondence to Emme C. K. Lin.

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Zhang, M.C., Wu, J., Ardlie, K.G. et al. Evaluation of clinical colon carcinoma using activity-based proteomic profiling. Clin Proteom 1, 301–311 (2004) doi:10.1385/CP:1:3-4:301

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Key Words

  • Tumor
  • activity
  • proteomics
  • colorectal adenocarcinoma
  • serine hydrolase