Startseite Recombinant kallikrein expression: site-specific integration for hK6 production in human cells
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Recombinant kallikrein expression: site-specific integration for hK6 production in human cells

  • Nathalie Heuzé-Vourc'h , Mireille Aïnciburu , Chris Planque , Michèle Brillard-Bourdet , Catherine Ott , Colette Jolivet-Reynaud und Yves Courty
Veröffentlicht/Copyright: 26. Juni 2006
Biological Chemistry
Aus der Zeitschrift Band 387 Heft 6

Abstract

Kallikreins have been implicated in carcinogenesis and are promising biomarkers for the diagnosis and follow-up of various cancers. To evaluate the functions and clinical interest of kallikreins, it is important to be able to produce them as recombinant proteins. Here we summarize the various strategies used to produce kallikreins, emphasizing their advantages and limitations. We also describe an approach to achieve high-level production of kallikreins, such as hK6, with correct folding and activity, combining an expression system with targeted transgene integration and an efficient cultivation device to increase yield, the CELLine bioreactor. This novel method for recombinant kallikrein production will be useful to study their bio-pathological functions and to develop anti-bodies.

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References

Angermann, A., Bergmann, C., and Appelhans, H. (1989). Cloning and expression of human salivary-gland kallikrein in Escherichia coli. Biochem. J.262, 787–793.10.1042/bj2620787Suche in Google Scholar

Angermann, A., Rahn, H.P., Hektor, T., Fertig, G., and Kemme, M. (1992). Purification and characterization of human salivary-gland prokallikrein from recombinant baculovirus-infected insect cells. Eur. J. Biochem.206, 225–233.10.1111/j.1432-1033.1992.tb16920.xSuche in Google Scholar

Baneyx, F. (1999). Recombinant protein expression in Escherichia coli. Curr. Opin. Biotechnol.10, 411–421.10.1016/S0958-1669(99)00003-8Suche in Google Scholar

Barnes, L.M., Bentley, C.M., and Dickson, A.J. (2003). Stability of protein production from recombinant mammalian cells. Biotechnol. Bioeng.81, 631–639.10.1002/bit.10517Suche in Google Scholar

Bayes, A., Tsetsenis, T., Ventura, S., Vendrell, J., Aviles, F.X., and Sotiropoulou, G. (2004). Human kallikrein 6 activity is regulated via an autoproteolytic mechanism of activation/inactivation. Biol. Chem.385, 517–524.10.1515/BC.2004.061Suche in Google Scholar

Bernett, M.J., Blaber, S.I., Scarisbrick, I.A., Dhanarajan, P., Thompson, S.M., and Blaber, M. (2002). Crystal structure and biochemical characterization of human kallikrein 6 reveals that a trypsin-like kallikrein is expressed in the central nervous system. J. Biol. Chem.277, 24562–24570.10.1074/jbc.M202392200Suche in Google Scholar

Borgoño, C.A. and Diamandis, E.P. (2004). The emerging roles of human tissue kallikreins in cancer. Nat. Rev. Cancer4, 876–890.10.1038/nrc1474Suche in Google Scholar

Borgoño, C.A., Grass, L., Soosaipillai, A., Yousef, G.M., Petraki, C.D., Howarth, D.H.C., Fracchioli, S., Katsaros, D., and Diamandis, E.P. (2003). Human kallikrein 14: a new potential biomarker for ovarian and breast cancer. Cancer Res.63, 9032–9041.Suche in Google Scholar

Brattsand, M. and Egelrud, T. (1999). Purification, molecular cloning, and expression of a human stratum corneum trypsin-like serine protease with possible function in desquamation. J. Biol. Chem.274, 30033–30040.10.1074/jbc.274.42.30033Suche in Google Scholar

Clark, E. (1998). Refolding of recombinant proteins. Curr. Opin. Biotechnol.9, 157–163.10.1016/S0958-1669(98)80109-2Suche in Google Scholar

Daly, R. and Hearn, M.T.W. (2005). Expression of heterologous proteins in Pichia pastoris: a useful experimental tool in protein engineering and production. J. Mol. Recognit.18, 119–138.10.1002/jmr.687Suche in Google Scholar PubMed

Eerola, R., Piironen, T., Pettersson, K., Lovgren, J., Vehniainen, M., Lilja, H., Dowell, B., Lovgren, T., and Karp, M. (1997). Immunoreactivity of recombinant human glandular kallikrein using monoclonal antibodies raised against prostate-specific antigen. Prostate31, 84–90.10.1002/(SICI)1097-0045(19970501)31:2<84::AID-PROS2>3.0.CO;2-HSuche in Google Scholar

Ekholm, I.E., Brattsand, M., and Egelrud, T. (2000). Stratum corneum tryptic enzyme in normal epidermis: a missing link in the desquamation process?J. Invest. Dermatol.114, 56–63.10.1046/j.1523-1747.2000.00820.xSuche in Google Scholar

Fortier, A.H., Holaday, J.W., Liang, H., Dey, C., Grella, D.K., Holland-Linn, J., Vu, H., Plum, S.M., and Nelson, B.J. (2003). Recombinant prostate specific antigen inhibits angiogenesis in vitro and in vivo. Prostate56, 212–219.10.1002/pros.10256Suche in Google Scholar

Habeck, L.L., Belagaje, R.M., Becker, G.W., Hale, J.E., Churgay, L.M., Ulmer, M., Yang, X.Y., Shackelford, K.A., Richardson, J.M., Johnson, M.G., and Mendelsohn, L.G. (2001). Expression, purification, and characterization of active recombinant prostate-specific antigen in Pichia pastoris (yeast). Prostate46, 298–306.10.1002/1097-0045(20010301)46:4<298::AID-PROS1036>3.0.CO;2-JSuche in Google Scholar

Hansson, L., Stromqvist, M., Backman, A., Wallbrandt, P., Carlstein, A., and Egelrud, T. (1994). Cloning, expression, and characterization of stratum corneum chymotryptic enzyme. A skin-specific human serine proteinase. J. Biol. Chem.269, 19420–19426.Suche in Google Scholar

Huang, X., Knoell, C.T., Frey, G., Hazegh-Azam, M., Tashjian, A.H.J., Hedstrom, L., Abeles, R.H., and Timasheff, S.N. (2001). Modulation of recombinant human prostate-specific antigen: activation by Hofmeister salts and inhibition by azapeptides. Appendix: thermodynamic interpretation of the activation by concentrated salts. Biochemistry40, 11734–11741.Suche in Google Scholar

Kapadia, C., Chang, A., Sotiropoulou, G., Yousef, G.M., Grass, L., Soosaipillai, A., Xing, X., Howarth, D.H.C., and Diamandis, E.P. (2003). Human kallikrein 13: production and purification of recombinant protein and monoclonal and polyclonal antibodies, and development of a sensitive and specific immunofluorometric assay. Clin. Chem.49, 77–86.10.1373/49.1.77Suche in Google Scholar

Karr, J.F., Kantor, J.A., Hand, P.H., Eggensperger, D.L., and Schlom, J. (1995). The presence of prostate-specific antigen-related genes in primates and the expression of recombinant human prostate-specific antigen in a transfected murine cell line. Cancer Res.55, 2455–2462.Suche in Google Scholar

Katz, B.A., Liu, B., Barnes, M., and Springman, E.B. (1998). Crystal structure of recombinant human tissue kallikrein at 2.0 Å resolution. Protein Sci.7, 875–885.10.1002/pro.5560070405Suche in Google Scholar

Kishi, T., Kato, M., Shimizu, T., Kato, K., Matsumoto, K., Yoshida, S., Shiosaka, S., and Hakoshima, T. (1997). Crystallization and preliminary X-ray analysis of neuropsin, a serine protease expressed in the limbic system of mouse brain. J. Struct. Biol.118, 248–251.10.1006/jsbi.1997.3862Suche in Google Scholar

Kishi, T., Grass, L., Soosaipillai, A., Shimizu-Okabe, C., and Diamandis, E.P. (2003). Human kallikrein 8: immunoassay development and identification in tissue extracts and biological fluids. Clin. Chem.49, 87–96.10.1373/49.1.87Suche in Google Scholar

Kishi, T., Soosaipillai, A., Grass, L., Little, S.P., Johnstone, E.M., and Diamandis, E.P. (2004). Development of an immunofluorometric assay and quantification of human kallikrein 7 in tissue extracts and biological fluids. Clin. Chem.50, 709–716.10.1373/clinchem.2003.029538Suche in Google Scholar

Kumar, A., Goel, A.S., Hill, T.M., Mikolajczyk, S.D., Millar, L.S., Kuus-Reichel, K., and Saedi, M.S. (1996). Expression of human glandular kallikrein, hK2, in mammalian cells. Cancer Res.56, 5397–5402.Suche in Google Scholar

Kumar, A., Goel, A.S., Payne, J.K., Evans, C., Mikolajczyk, S.D., Kuus-Reichel, K., and Saedi, M.S. (1999). Large-scale propagation of recombinant adherent cells that secrete a stable form of human glandular kallikrein, hK2. Protein Expr. Purif.15, 62–68.10.1006/prep.1998.0998Suche in Google Scholar

Kurkela, R., Herrala, A., Henttu, P., Nai, H., and Vihko, P. (1995). Expression of active, secreted human prostate-specific antigen by recombinant baculovirus-infected insect cells on a pilot-scale. Biotechnology13, 1230–1234.10.1038/nbt1195-1230Suche in Google Scholar

Laxmikanthan, G., Blaber, S.I., Bernett, M.J., Scarisbrick, I.A., Juliano, M.A., and Blaber, M. (2005). 1.70 Å X-ray structure of human apo kallikrein 1: structural changes upon peptide inhibitor/substrate binding. Proteins58, 802–814.Suche in Google Scholar

Lovgren, J., Piironen, T., Overmo, C., Dowell, B., Karp, M., Pettersson, K., Lilja, H., and Lundwall, A. (1995). Production of recombinant PSA and HK2 and analysis of their immunologic cross-reactivity. Biochem. Biophys. Res. Commun.213, 888–895.10.1006/bbrc.1995.2212Suche in Google Scholar

Lovgren, J., Rajakoski, K., Karp, M., and Lilja, H. (1997). Activation of the zymogen form of prostate-specific antigen by human glandular kallikrein 2. Biochem. Biophys. Res. Commun.238, 549–555.10.1006/bbrc.1997.7333Suche in Google Scholar

Lovgren, J., Tian, S., Lundwall, A., Karp, M., and Lilja, H. (1999). Production and activation of recombinant hK2 with propeptide mutations resulting in high expression levels. Eur. J. Biochem.266, 1050–1055.10.1046/j.1432-1327.1999.00946.xSuche in Google Scholar

Lu, H.S., Hsu, Y.R., Lu, L.I., Ruff, D., Lyons, D., and Lin, F.K. (1996). Isolation and characterization of human tissue kallikrein produced in Escherichia coli: biochemical comparison to the enzymatically inactive prokallikrein and methionyl kallikrein. Protein Expr. Purif.8, 215–226.10.1006/prep.1996.0094Suche in Google Scholar

Lundstrom, K. (2003). Semliki Forest virus vectors for rapid and high-level expression of integral membrane proteins. Biochim. Biophys. Acta1610, 90–96.10.1016/S0005-2736(02)00721-6Suche in Google Scholar

Luo, L.Y., Grass, L., Howarth, D.J., Thibault, P., Ong, H., and Diamandis, E.P. (2001). Immunofluorometric assay of human kallikrein 10 and its identification in biological fluids and tissues. Clin. Chem.47, 237–246.10.1093/clinchem/47.2.237Suche in Google Scholar

Magklara, A., Mellati, A.A., Wasney, G.A., Little, S.P., Sotiropoulou, G., Becker, G.W., and Diamandis, E.P. (2003). Characterization of the enzymatic activity of human kallikrein 6: autoactivation, substrate specificity, and regulation by inhibitors. Biochem. Biophys. Res. Commun.307, 948–955.10.1016/S0006-291X(03)01271-3Suche in Google Scholar

Matsumura, M., Bhatt, A.S., Andress, D., Clegg, N., Takayama, T.K., Craik, C.S., and Nelson, P.S. (2005). Substrates of the prostate-specific serine protease prostase/KLK4 defined by positional-scanning peptide libraries. Prostate62, 1–13.10.1002/pros.20101Suche in Google Scholar PubMed

Mergulhao, F.J.M., Summers, D.K., and Monteiro, G.A. (2005). Recombinant protein secretion in Escherichia coli. Biotechnol. Adv.23, 177–202.10.1016/j.biotechadv.2004.11.003Suche in Google Scholar PubMed

Michael, I.P., Sotiropoulou, G., Pampalakis, G., Magklara, A., Ghosh, M., Wasney, G., and Diamandis, E.P. (2005). Biochemical and enzymatic characterization of human kallikrein 5 (hK5), a novel serine protease potentially involved in cancer progression. J. Biol. Chem.280, 14628–14635.10.1074/jbc.M408132200Suche in Google Scholar

Mitsui, S., Yamada, T., Okui, A., Kominami, K., Uemura, H., and Yamaguchi, N. (2000). A novel isoform of a kallikrein-like protease, TLSP/hippostasin, (PRSS20), is expressed in the human brain and prostate. Biochem. Biophys. Res. Commun.272, 205–211.10.1006/bbrc.2000.2761Suche in Google Scholar

Obiezu, C.V., Soosaipillai, A., Jung, K., Stephan, C., Scorilas, A., Howarth, D.H.C., and Diamandis, E.P. (2002). Detection of human kallikrein 4 in healthy and cancerous prostatic tissues by immunofluorometry and immunohistochemistry. Clin. Chem.48, 1232–1240.10.1093/clinchem/48.8.1232Suche in Google Scholar

Saedi, M.S., Cass, M.M., Goel, A.S., Grauer, L., Hogen, K.L., Okaneya, T., Griffin, B.Y., Klee, G.G., Young, C.Y., and Tindall, D.J. (1995). Overexpression of a human prostate-specific glandular kallikrein, hK2, in E. coli and generation of antibodies. Mol. Cell. Endocrinol.109, 237–241.10.1016/0303-7207(95)03508-5Suche in Google Scholar

Saedi, M.S., Zhu, Z., Marker, K., Liu, R.S., Carpenter, P.M., Rittenhouse, H., and Mikolajczyk, S.D. (2001). Human kallikrein 2 (hK2), but not prostate-specific antigen (PSA), rapidly complexes with protease inhibitor 6 (PI-6) released from prostate carcinoma cells. Int. J. Cancer94, 558–563.10.1002/ijc.1501Suche in Google Scholar PubMed

Schmidt, F.R. (2004). Recombinant expression systems in the pharmaceutical industry. Appl. Microbiol. Biotechnol.65, 363–372.10.1007/s00253-004-1656-9Suche in Google Scholar PubMed

Shinmura, K., Tao, H., Yamada, H., Kataoka, H., Sanjar, R., Wang, J., Yoshimura, K., and Sugimura, H. (2004). Splice-site genetic polymorphism of the human kallikrein 12 (KLK12) gene correlates with no substantial expression of KLK12 protein having serine protease activity. Hum. Mutat.24, 273–274.10.1002/humu.9270Suche in Google Scholar PubMed

Takayama, T.K., Fujikawa, K., and Davie, E.W. (1997). Characterization of the precursor of prostate-specific antigen. Activation by trypsin and by human glandular kallikrein. J. Biol. Chem.272, 21582–21588.10.1074/jbc.272.34.21582Suche in Google Scholar PubMed

Takayama, T.K., Carter, C.A., and Deng, T. (2001a). Activation of prostate-specific antigen precursor (pro-PSA) by prostin, a novel human prostatic serine protease identified by degenerate PCR. Biochemistry40, 1679–1687.10.1021/bi002129rSuche in Google Scholar PubMed

Takayama, T.K., McMullen, B.A., Nelson, P.S., Matsumura, M., and Fujikawa, K. (2001b). Characterization of hK4 (prostase), a prostate-specific serine protease: activation of the precursor of prostate specific antigen (pro-PSA) and single-chain urokinase-type plasminogen activator and degradation of prostatic acid phosphatase. Biochemistry40, 15341–15348.10.1021/bi015775eSuche in Google Scholar PubMed

Thyagarajan, B. and Calos, M.P. (2005). Site-specific integration for high-level protein production in mammalian cells. Methods Mol. Biol.308, 99–106.10.1385/1-59259-922-2:099Suche in Google Scholar

Wurm, F.M. (2004). Production of recombinant protein therapeutics in cultivated mammalian cells. Nat. Biotechnol.22, 1393–1398.10.1038/nbt1026Suche in Google Scholar

Yousef, G.M. and Diamandis, E.P. (2001). The new human tissue kallikrein gene family: structure, function, and association to disease. Endocr. Rev.22, 184–204.Suche in Google Scholar

Yousef, G.M., Kapadia, C., Polymeris, M., Borgono, C., Hutchinson, S., Wasney, G.A., Soosaipillai, A., and Diamandis, E.P. (2003a). The human kallikrein protein 5 (hK5) is enzymatically active, glycosylated and forms complexes with two protease inhibitors in ovarian cancer fluids. Biochim. Biophys. Acta1628, 88–96.10.1016/S0167-4781(03)00116-7Suche in Google Scholar

Yousef, G.M., Polymeris, M., Grass, L., Soosaipillai, A., Chan, P., Scorilas, A., Borgono, C., Harbeck, N., Schmalfeldt, B., Dorn, J., et al. (2003b). Human kallikrein 5: a potential novel serum biomarker for breast and ovarian cancer. Cancer Res.63, 3958–3965.Suche in Google Scholar

Published Online: 2006-06-26
Published in Print: 2006-06-01

©2006 by Walter de Gruyter Berlin New York

Artikel in diesem Heft

  1. The First International Symposium on Kallikreins
  2. A comprehensive nomenclature for serine proteases with homology to tissue kallikreins
  3. The kallikrein world: an update on the human tissue kallikreins
  4. Cellular distribution of human tissue kallikreins: immunohistochemical localization
  5. The tissue kallikrein-kinin system protects against cardiovascular and renal diseases and ischemic stroke independently of blood pressure reduction
  6. Proteinase-mediated cell signalling: targeting proteinase-activated receptors (PARs) by kallikreins and more
  7. Recombinant kallikrein expression: site-specific integration for hK6 production in human cells
  8. Kallikrein-related peptidase (KLK) family mRNA variants and protein isoforms in hormone-related cancers: do they have a function?
  9. The role of kallikrein-related peptidases in prostate cancer: potential involvement in an epithelial to mesenchymal transition
  10. Human kallikrein 10, a predictive marker for breast cancer
  11. Activation and enzymatic characterization of recombinant human kallikrein 8
  12. Human tissue kallikrein 9: production of recombinant proteins and specific antibodies
  13. The human kallikrein 10 promoter contains a functional retinoid response element
  14. Human kallikrein 4: enzymatic activity, inhibition, and degradation of extracellular matrix proteins
  15. Kallikrein-related peptidase 14 may be a major contributor to trypsin-like proteolytic activity in human stratum corneum
  16. A sensitive proximity ligation assay for active PSA
  17. Multiple mechanisms underlie the aberrant expression of the human kallikrein 6 gene in breast cancer
  18. Expression of the human kallikrein genes 10 (KLK10) and 11 (KLK11) in cancerous and non-cancerous lung tissues
  19. mRNA expression analysis of human kallikrein 11 (KLK11) may be useful in the discrimination of benign prostatic hyperplasia from prostate cancer after needle prostate biopsy
  20. The epigenetic basis for the aberrant expression of kallikreins in human cancers
  21. Improved prostate cancer detection with a human kallikrein 11 and percentage free PSA-based artificial neural network
  22. Overexpression of the human tissue kallikrein genes KLK4, 5, 6, and 7 increases the malignant phenotype of ovarian cancer cells
  23. Inhibition profiles of human tissue kallikreins by serine protease inhibitors
  24. Kallikrein-mediated cell signalling: targeting proteinase-activated receptors (PARs)
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