Browse > Article

Oligomeric Structure of the ATP-dependent Protease La (Lon) of Escherichia coli  

Park, Seong-Cheol (Division of Applied Life Science, Gyeongsang National University)
Jia, Baolei (Division of Applied Life Science, Gyeongsang National University)
Yang, Jae-Kyung (Faculty of Forest Science, Gyeongsang National University)
Le Van, Duyet (Division of Applied Life Science, Gyeongsang National University)
Shao, Yong Gi (Division of Applied Life Science, Gyeongsang National University)
Han, Sang Woo (Department of Chemistry, Gyeongsang National University)
Jeon, Young-Joo (NRL of Protein Biochemistry, School of Biological Science, Seoul National University)
Chung, Chin Ha (NRL of Protein Biochemistry, School of Biological Science, Seoul National University)
Cheong, Gang-Won (Division of Applied Life Science, Gyeongsang National University)
Publication Information
Molecules and Cells / v.21, no.1, 2006 , pp. 129-134 More about this Journal
Abstract
Lon, also known as protease La, belongs to a class of ATP-dependent serine protease. It plays an essential role in degradation of abnormal proteins and of certain short-lived regulatory proteins, and is thought to possess a Ser-Lys catalytic dyad. To examine the structural organization of Lon, we performed an electron microscope analysis. The averaged images of Lon with end-on orientation revealed a six-membered, ring-shaped structure with a central cavity. The side-on view showed a two-layered structure with an equal distribution of mass across the equatorial plane of the complex. Since a Lon subunit possesses two large regions containing nucleotide binding and proteolytic domains, each layer of the Lon hexamer appears to consist of the side projections of one of the major domains arranged in a ring. Lon showed a strong tendency to form hexamers in the presence of $Mg^{2+}$, but dissociated into monomers and/or dimers in its absence. Moreover, $Mg^{2+}$-dependent hexamer formation was independent of ATP. These results indicate that Lon has a hexameric ring-shaped structure with a central cavity, and that the establishment of this configuration requires $Mg^{2+}$, but not ATP.
Keywords
ATP-dependent Protease La; Electron Microscopy; Lon; Oligomerization;
Citations & Related Records
Times Cited By KSCI : 1  (Citation Analysis)
Times Cited By Web Of Science : 21  (Related Records In Web of Science)
연도 인용수 순위
1 Birghan, C., Mundt, E., and Gorbalenya, A. E. (2000) A noncanonical Lon proteinase lacking the ATPase domain employs the Ser-Lys catalytic dyad to exercise broad control over the life cycle of a double-stranded RNA virus. EMBO J. 19, 114-123   DOI   ScienceOn
2 Chung, C. H. and Goldberg, A. L. (1981) The product of the lon (capR) gene in Escherichia coli is the ATP-dependent protease, protease La. Proc. Natl. Acad. Sci. USA 78, 4931−4935
3 Chung, C. H., Yoo, S. J., Seol, J. H., and Kang, M. S. (1997) Characterization of energy-dependent proteases in bacteria. Biochem. Biophys. Res. Commun. 241, 613-616   DOI   ScienceOn
4 Goldberg, A. L., Moerschell, R. P., Chung, C. H., and Maurizi, M. R. (1994) ATP-dependent protease La (lon) from Escherichia coli. Methods Enzymol. 244, 350-375   DOI
5 Gottesman, S. (2003) Proteolysis in bacterial regulatory circuits. Annu. Rev. Cell Dev. Biol. 19, 565−587   DOI   ScienceOn
6 Grimaud, R., Kessel, M., Beuron, F., Steven, A. C., and Maurizi, M. R. (1998) Enzymatic and structural similarities between the Escherichia coli ATP-dependent proteases, ClpXP and ClpAP. J. Biol. Chem. 273, 12476-12481   DOI   ScienceOn
7 Hegerl, R. (1996) The EM program package: a platform for image processing in biological electron microscopy. J. Struct. Biol. 116, 30−34
8 Kim, K. I., Cheong, G.-W., Park, S. C., Ha, J. S., Woo, K. M., et al. (2000) Heptameric ring structure of the heat-shock protein ClpB, a protein-activated ATPase in Escherichia coli. J. Mol. Biol. 303, 655-666   DOI   ScienceOn
9 Oh, J. Y., Eun, Y. M., Yoo, S. J., Seol, J. H., Seong, I. S., et al. (1998) LonR9 carrying a single $Glu^{614}$ to Lys mutation inhibits the ATP-dependent protease La (Lon) by forming mixed oligomeric complexes. Biochem. Biophys. Res. Commun. 250, 32-35   DOI   ScienceOn
10 Rudyak, S. G., Brenowitz, M., and Shrader, T. E. (2001) $Mg^{2+}$-linked oligomerization modulates the catalytic activity of the Lon (La) protease from Mycobacterium smegmatis. Biochemistry 40, 9317-9323   DOI   ScienceOn
11 Schirmer, E. C., Glover, J. R., Singer, M. A., and Lindquist, S. (1996) HSP100/Clp proteins: a common mechanism explains diverse functions. Trends Biochem. Sci. 21, 289−296
12 Vasilyeva, O. V., Kolygo, K. B., Leonova, Y. F., Potapenko, N. A., and Ovchinnikova, T. V. (2002) Domain structure and ATP-induced conformational changes in Escherichia coli protease Lon revealed by limited proteolysis and autolysis. FEBS Lett. 526, 66-70   DOI
13 Woo, K. M., Kim, K. I., Goldberg, A. L., Ha, D. B., and Chung, C. H. (1992) The heat-shock protein ClpB in Escherichia coli is a protein-activated ATPase. J. Biol. Chem. 267, 20429-20434
14 Yoo, S. J., Seol, J. H., Seong, I. S., Kang, M. S., and Chung, C. H. (1997) ATP binding, but not its hydrolysis, is required for assembly and proteolytic activity of the HslVU protease in Escherichia coli. Biochem. Biophys. Res. Commun. 238, 581−585   DOI   ScienceOn
15 Zolkiewski, M., Kessel, M., Ginsburg, A., and Maurizi, M. R. (1999) Nucleotide-dependent oligomerization of ClpB from Escherichia coli. Protein Sci. 8, 1899-1903   DOI   ScienceOn
16 Wang, J., Song, J. J., Im, Y. J., Rho, S. H., Seong, I. S., et al. (2001) Crystal structures of the HslVU peptidase-ATPase complex reveal an ATP-dependent proteolysis mechanism. Structure 9, 177-184   DOI   ScienceOn
17 Chung, C. H. (1993) Proteases in Escherichia coli. Science 262, 372−374   DOI
18 Maurizi, M. R., Singh, S. K., Thompson, M. W., Kessel, M., and Ginsburg, A. (1998) Molecular properties of ClpAP protease of Escherichia coli: ATP-dependent association of ClpA and ClpP. Biochemistry 37, 7778-7786   DOI   ScienceOn
19 Wang, J., Hartling, J. A., and Flanagan, J. M. (1997) The structure of ClpP at 2.3 ${\AA}$ resolution suggests a model for ATPdependent proteolysis. Cell 91, 447−456   DOI   ScienceOn
20 Benaroudj, N., Zwickl, P., Seemüller, E., Baumeister, W., and Goldberg, A. L. (2003) ATP hydrolysis by the proteasome regulatory complex PAN serves multiple functions in protein degradation. Mol. Cell 11, 69−78   DOI   ScienceOn
21 Sousa, M. C., Trame, C. B., Tsuruta, S., Wilbanks, S. M., Reddy, R. S., et al. (2000) Crystal and solution structures of an HslUV protease-chaperone complex. Cell 103, 633-643   DOI   ScienceOn
22 Kang, M. S., Kim, S. R., Kwack, P., Lim, B. K., Ahn, S. W., et al. (2003) Molecular architecture of the ATP-dependent CodWX protease having an N-terminal serine active site. EMBO J. 22, 2893-2902   DOI   ScienceOn
23 Baumeister, W., Walz, J., Zuhl, F., and Seemüller, E. (1998b) The proteasome: paradigm of a self-compartmentalizing protease. Cell 92, 367-380   DOI   ScienceOn
24 Saxton, W. O., Pitt, T. J., and Horner, M. (1979) Digital image processing: the Semper system. Ultramicroscopy 4, 343−354   DOI   ScienceOn
25 Hoskins, J. R., Park, M., Maurizi, M. R., and Wickner, S. (1998) The role of the ClpA chaperone in proteolysis by ClpAP. Proc. Natl. Acad. Sci. USA 95, 12135−12140
26 Gottesman, S. (1996) Proteases and their targets in Escherichia coli. Annu. Rev. Genet. 30, 465−506   DOI   ScienceOn
27 Chandu, D. and Nandi, D. (2004) Comparative genomics and functional roles of the ATP-dependent proteases Lon and Clp during cytosolic protein degradation. Res. Microbiol. 155, 710-719
28 Chin, D. T., Goff, S. A., Webster, T., Smith, T., and Goldberg, A. L. (1988) Sequence of the lon gene in Escherichia coli. A heat-shock gene which encodes the ATP-dependent protease La. J. Biol. Chem. 263, 11718-11728
29 van Heel, M. and Frank, J. (1981) Use of multivariate statistics in analyzing the images of biological macromolecules. Ultramicroscopy 6, 187−194
30 Coux, O., Tanaka, K., and Goldberg, A. L. (1996) Structure and functions of the 20S and 26S proteasomes. Annu. Rev. Biochem. 65, 801−847   DOI   ScienceOn
31 Marco, S., Urena, D., Carrascosa, J. L., Waldmann, T., Peter, J., et al. (1994) The molecular chaperone TF55: assessment of symmetry. FEBS Lett. 341, 152−155   DOI
32 Rohrwild, M., Pfeifer, G., Santarius, U., Müller, S. A., Huang, H. C., et al. (1997) The ATP-dependent HslVU protease from Escherichia coli is a four-ring structure resembling the proteasome. Nat. Struct. Biol. 4, 133-139   DOI   ScienceOn
33 Stahlberg, H., Kutejová, E., Suda, K., Wolpensinger, B., Lustig, A., et al. (1999) Mitochondrial Lon of Saccharomyces cerevisiae is a ring-shaped protease with seven flexible subunits. Proc. Natl. Acad. Sci. USA 96, 6787−6790
34 Chung, C. H., Waxman, L., and Goldberg, A. L. (1983) Studies of the protein encoded by the lon mutation, capR9, in Escherichia coli. A labile form of the ATP-dependent protease La that inhibits the wild type protease. J. Biol. Chem. 258, 215-221
35 Yoo, S. J., Seol, J. H., Shin, D. H., Rohrwild, M., Kang, M. S., et al. (1996) Purification and characterization of the heat shock proteins HslV and HslU that form a new ATPdependent protease in Escherichia coli. J. Biol. Chem. 271, 14035−14040
36 Ishikawa, T., Maurizi, M. R., Belnap, D., and Steven, A. C. (2000) Docking of components in a bacterial complex. Nature 408, 667−668   DOI
37 Goldberg, A. L. (1992) The mechanism and function of ATPdependent protease in bacterial and animal cells. Eur. J. Biochem. 203, 9−23
38 Botos, I., Melnikov, E. E., Cherry, S., Tropea, J. E., Khalatova, A. G., et al. (2004) The catalytic domain of Escherichia coli Lon protease has a unique fold and a Ser-Lys dyad in the active site. J. Biol. Chem. 279, 8140-8148   DOI   ScienceOn
39 Beuron, F., Maurizi, M. R., Belnap, D. M., Kocsis, E., Booy, F. P., et al. (1998) At sixes and sevens: characterization of the symmetry mismatch of the ClpAP chaperone-assisted protease. J. Struct. Biol. 123, 248-259   DOI   ScienceOn
40 Kessel, M., Wu, W., Gottesman, S., Kocsis, E., Steven, A. C., et al. (1996) Six-fold rotational symmetry of ClpQ, the E. coli homolog of the 20S proteasome and its ATP-dependent activator, ClpY. FEBS Lett. 398, 274-278   DOI
41 Baumeister, W., Dahlmann, B., Hegerl, R., Kopp, F., Luehn, L., et al. (1988a) Electron microscopy and image analysis of the multicatalytic proteinase. FEBS Lett. 241, 239−245   DOI
42 Durr, R. (1991) Displacement field analysis: calculation of distortion measures from displacement maps. Ultramicroscopy 38, 135−141   DOI   ScienceOn