N-terminal truncation circumvents proteolytic degradation of the human HtrA2/Omi serine protease in Escherichia coli: Rapid purification of a proteolytically active HtrA2/Omi

Protein Expression and Purification

Volumn 33, Issue 2, 2004, Pages 200-208

  Seong, Young Mo ^c   Park, Hyo Jin ^c   Seong, Geun Hye ^c   Choi, Ju Youn ^a,d   Yoon, Sung Joo Kim ^a,b   Min, Byung Re ^d   Kang, Seongman ^c   Rhim, Hyangshuk ^a,b  

^a The Catholic University of Korea   (South Korea)

^b The Catholic University of Korea   (South Korea)

^c Korea University   (South Korea)

^d Sangmyung University   (South Korea)

Author keywords

hHtrA2 Omi; mHtrA2 Omi; Mitochondrial serine protease; pGEX expression system; PRAXXTXXTP motif; Proteolytic degradation

Indexed keywords

BACTERIA (MICROORGANISMS); ESCHERICHIA COLI;

EID: 0842331856     PISSN: 10465928     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.pep.2003.10.002     Document Type: Article

References (22)

1. J.Y. Choi, Y.M. Sung, H.J. Park, E.H. Hur, S.J. Lee, C. Hahn, B.R. Min, I.K. Kim, S. Kang, H. Rhim, Rapid purification and analysis of alpha-synuclein proteins: C-terminal truncation promotes the conversion of alpha-synuclein into a protease-sensitive form in Escherichia coli, Biotechnol. Appl. Biochem. 36 (2002) 33-40.
2. L. Cilenti, Y. Lee, S. Hess, S. Srinivasula, K.M. Park, D. Junqueira, H. Davis, J.V. Bonventre, E.S. Alnemri, A.S. Zervos, Characterization of a novel and specific inhibitor for the pro-apoptotic protease Omi/HtrA2, J. Biol. Chem. 278 (2003) 11489-11494.
3. T. Clausen, C. Southan, M. Ehrmann, The HtrA family of proteases: implications for protein composition and cell fate, Mol. Cell 10 (2002) 443-455.
4. L. Faccio, C. Fusco, A. Chen, S. Martinotti, J.V. Bonventre, A.S. Zervos, Characterization of a novel human serine protease that has extensive homology to bacterial heat shock endoprotease HtrA and is regulated by kidney ischemia, J. Biol. Chem. 275 (2000) 2581-2588.
5. C.W. Gray, R.V. Ward, E. Karran, S. Turconi, A. Rowles, D. Viglienghi, C. Southan, A. Barton, K.G. Fantom, A. West, J. Savopoulos, N.J. Hassan, H. Clinkenbeard, C. Hanning, B. Amegadzie, J.B. Davis, C. Dingwall, G.P. Livi, C.L. Creasy, Characterization of human HtrA2, a novel serine protease involved in the mammalian cellular stress response, Eur. J. Biochem. 267 (2000) 5699-5710.
6. J. Grodberg, J.J. Dunn, ompT encodes the Escherichia coli outer membrane protease that cleaves T7 RNA polymerase during purification, J. Bacteriol. 170 (1988) 1245-1253.
7. R. Hegde, S.M. Srinivasula, Z. Zhang, R. Wassell, R. Mukattash, L. Cilenti, G. DuBois, Y. Lazebnik, A.S. Zervos, T. Fernandes-Alnemri, E.S. Alnemri, Identification of Omi/HtrA2 as a mitochondrial apoptotic serine protease that disrupts inhibitor of apoptosis protein-caspase interaction, J. Biol. Chem. 277 (2002) 432-438.
8. M. Leverkus, M.R. Sprick, T. Wachter, T. Mengling, B. Baumann, E. Serfling, E.B. Brocker, M. Goebeler, M. Neumann, H. Walczak, Proteasome inhibition results in TRAIL sensitization of primary keratinocytes by removing the resistance-mediating block of effector caspase maturation, Mol. Cell. Biol. 23 (2003) 777-790.
9. W. Li, S.M. Srinivasula, J. Chai, P. Li, J.W. Wu, Z. Zhang, E.S. Alnemri, Y. Shi, Structural insights into the pro-apoptotic function of mitochondrial serine protease HtrA2/Omi, Nat. Struct. Biol. 9 (2002) 436-441.
10. L.M. Martins, I. Iaccarino, T. Tenev, S. Gschmeissner, N.F. Totty, N.R. Lemoine, J. Savopoulos, C.W. Gray, C.L. Creasy, C. Dingwall, J. Downward, The serine protease Omi/HtrA2 regulates apoptosis by binding XIAP through a reaper-like motif, J. Biol. Chem. 277 (2002) 439-444.
11. M. Murby, M. Uhlen, S. Stahl, Upstream strategies to minimize proteolytic degradation upon recombinant production in Escherichia coli, Protein Expr. Purif. 7 (1996) 129-136.
12. W.R. Pearson, T. Wood, Z. Zhang, W. Miller, Comparison of DNA sequences with protein sequences, Genomics 46 (1997) 24-36.
13. L. Ravagnan, T. Roumier, G. Kroemer, Mitochondria, the killer organelles and their weapons, J. Cell Physiol. 192 (2002) 131-137.
14. H. Rhim, F. Chan, C.O. Echetebu, A.P. Rice, Rapid purification of monomer HIV-2 Tat protein expressed in Escherichia coli, Protein Expr. Purif. 4 (1993) 24-31.
15. H. Rhim, A.P. Rice, TAR RNA binding properties and relative transactivation activities of human immunodeficiency virus type 1 and 2 Tat proteins, J. Virol. 67 (1993) 1110-1121.
16. J.W. Savopoulos, P.S. Carter, S. Turconi, G.R. Pettman, E.H. Karran, C.W. Gray, R.V. Ward, O. Jenkins, C.L. Creasy, Expression purification and functional analysis of the human serine protease HtrA2, Protein Expr. Purif. 19 (2000) 227-234.
17. J. Silke, C.J. Hawkins, P.G. Ekert, J. Chew, C.L. Day, M. Pakusch, A.M. Verhagen, D.L. Vaux, The anti-apoptotic activity of XIAP is retained upon mutation of both the caspase 3- and caspase 9-interacting sites, J. Cell Biol. 157 (2002) 115-124.
18. Y. Suzuki, Y. Imai, H. Nakayama, K. Takahashi, K. Takio, R. Takahashi, A serine protease, HtrA2, is released from the mitochondria and interacts with XİAP, inducing cell death, Mol. Cell 8 (2001) 613-621.
19. G. van Loo, X. Saelens, M. van Gurp, M. MacFarlane, S.J. Martin, P. Vandenabeele, The role of mitochondrial factors in apoptosis: a Russian roulette with more than one bullet, Cell Death Differ. 9 (2002) 1031-1042.
20. G. van Loo, M. van Gurp, B. Depuydt, S.M. Srinivasula, I. Rodriguez, E.S. Alnemri, K. Gevaert, J. Vandekerckhove, W. Declercq, P. Vandenabeele, The serine protease Omi/HtrA2 is released from mitochondria during apoptosis. Omi interacts with caspase-inhibitor XIAP and induces enhanced caspase activity, Cell Death Differ. 9 (2002) 20-26.
21. A.M. Verhagen, J. Silke, P.G. Ekert, M. Pakusch, H. Kaufmann, L.M. Connolly, C.L. Day, A. Tikoo, R. Burke, C. Wrobel, R.L. Moritz, R.J. Simpson, D.L. Vaux, HtrA2 promotes cell death through its serine protease activity and its ability to antagonize inhibitor of apoptosis proteins, J. Biol. Chem. 277 (2002) 445-454.
22. S. Wittmann, P. Bali, S. Donapaty, R. Nimmanapalli, F. Guo, H. Yamaguchi, M. Huang, R. Jove, H.G. Wang, K. Bhalla, Flavopiridol down-regulates antiapoptotic proteins and sensitizes human breast cancer cells to epothilone B-induced apoptosis, Cancer Res. 63 (2003) 93-99.