메뉴 건너뛰기




Volumn 192, Issue 16, 2010, Pages 4172-4180

Structure and function of CinD (YtjD) of Lactococcus lactis, a copper-induced nitroreductase involved in defense against oxidative stress

Author keywords

[No Author keywords available]

Indexed keywords

2,6 DICHLOROPHENOLINDOPHENOL; 4 NITROQUINOLINE 1 OXIDE; BACTERIAL RNA; CADMIUM; CALCIUM; CIND PROTEIN; COPPER; COPPER SULFATE; FLAVOPROTEIN; HYDROGEN PEROXIDE; IRON; MANGANESE; MESSENGER RNA; NICKEL; NITROREDUCTASE; PARAQUAT; REDUCED NICOTINAMIDE ADENINE DINUCLEOTIDE; SILVER; UNCLASSIFIED DRUG; ZINC; BACTERIAL PROTEIN; CATALASE; NICOTINAMIDE ADENINE DINUCLEOTIDE; OXIDIZING AGENT;

EID: 77955963082     PISSN: 00219193     EISSN: 10985530     Source Type: Journal    
DOI: 10.1128/JB.00372-10     Document Type: Article
Times cited : (28)

References (49)
  • 1
    • 65549107862 scopus 로고    scopus 로고
    • Manganese import is a key element of the OxyR response to hydrogen peroxide in Escherichia coli
    • Anjem, A., S. Varghese, and J. A. Imlay. 2009. Manganese import is a key element of the OxyR response to hydrogen peroxide in Escherichia coli. Mol. Microbiol. 72:844-858.
    • (2009) Mol. Microbiol. , vol.72 , pp. 844-858
    • Anjem, A.1    Varghese, S.2    Imlay, J.A.3
  • 3
    • 34547749438 scopus 로고    scopus 로고
    • Copper induction of lactate oxidase of Lactococcus lactis: A novel metal stress response
    • Barré, O., F. Mourlane, and M. Solioz. 2007. Copper induction of lactate oxidase of Lactococcus lactis: a novel metal stress response. J. Bacteriol. 189:5947-5954.
    • (2007) J. Bacteriol. , vol.189 , pp. 5947-5954
    • Barré, O.1    Mourlane, F.2    Solioz, M.3
  • 4
    • 0035021812 scopus 로고    scopus 로고
    • The complete genome sequence of the lactic acid bacterium lactococcus lactis ssp. lactis IL1403
    • DOI 10.1101/gr.GR-1697R
    • Bolotin, A., P. Wincker, S. Mauger, O. Jaillon, K. Malarme, J. Weissenbach, S. D. Ehrlich, and A. Sorokin. 2001. The complete genome sequence of the lactic acid bacterium Lactococcus lactis ssp. lactis IL1403. Genome Res. 11:731-753. (Pubitemid 32447860)
    • (2001) Genome Research , vol.11 , Issue.5 , pp. 731-753
    • Bolotin, A.1    Wincker, P.2    Mauger, S.3    Jaillon, O.4    Malarme, K.5    Weissenbach, J.6    Ehrlich, S.D.7    Sorokin, A.8
  • 5
    • 0025892415 scopus 로고
    • Purification and characterization of an oxygen-insensitive NAD(P)H nitroreductase from Enterobacter cloacae
    • Bryant, C., and M. DeLuca. 1991. Purification and characterization of an oxygen-insensitive NAD(P)H nitroreductase from Enterobacter cloacae. J. Biol. Chem. 266:4119-4125. (Pubitemid 21909328)
    • (1991) Journal of Biological Chemistry , vol.266 , Issue.7 , pp. 4119-4125
    • Bryant, C.1    DeLuca, M.2
  • 6
    • 77952056332 scopus 로고    scopus 로고
    • Copper stress affects iron homeostasis by destabilizing iron-sulfur cluster formation in Bacillus subtilis
    • Chillappagari, S., A. Seubert, H. Trip, O. P. Kuipers, M. A. Marahiel, and M. Miethke. 2010. Copper stress affects iron homeostasis by destabilizing iron-sulfur cluster formation in Bacillus subtilis. J. Bacteriol. 192:2512-2524.
    • (2010) J. Bacteriol. , vol.192 , pp. 2512-2524
    • Chillappagari, S.1    Seubert, A.2    Trip, H.3    Kuipers, O.P.4    Marahiel, M.A.5    Miethke, M.6
  • 7
    • 39649108421 scopus 로고    scopus 로고
    • Crystal structure of a minimal nitroreductase, ydjA, from Escherichia coli K12 with and without FMN cofactor
    • Choi, J. W., J. Lee, K. Nishi, Y. S. Kim, C. H. Jung, and J. S. Kim. 2008. Crystal structure of a minimal nitroreductase, ydjA, from Escherichia coli K12 with and without FMN cofactor. J. Mol. Biol. 377:258-267.
    • (2008) J. Mol. Biol. , vol.377 , pp. 258-267
    • Choi, J.W.1    Lee, J.2    Nishi, K.3    Kim, Y.S.4    Jung, C.H.5    Kim, J.S.6
  • 8
  • 9
    • 34250702076 scopus 로고    scopus 로고
    • Efficient transformation of Lactococcus lactis IL1403 and generation of knock-out mutants by homologous recombination
    • Gerber, S. D., and M. Solioz. 2007. Efficient transformation of Lactococcus lactis IL1403 and generation of knock-out mutants by homologous recombination. J. Basic Microbiol. 47:281-286.
    • (2007) J. Basic Microbiol. , vol.47 , pp. 281-286
    • Gerber, S.D.1    Solioz, M.2
  • 10
    • 34248590170 scopus 로고    scopus 로고
    • Escherichia coli has multiple enzymes that attack TNT and release nitrogen for growth
    • Gonzalez-Perez, M. M., P. van Dillewijn, R. M. Wittich, and J. L. Ramos. 2007. Escherichia coli has multiple enzymes that attack TNT and release nitrogen for growth. Environ. Microbiol. 9:1535-1540.
    • (2007) Environ. Microbiol. , vol.9 , pp. 1535-1540
    • Gonzalez-Perez, M.M.1    Van Dillewijn, P.2    Wittich, R.M.3    Ramos, J.L.4
  • 11
    • 0037344041 scopus 로고    scopus 로고
    • Proteomic analysis of Lactococcus lactis, a lactic acid bacterium
    • Guillot, A., C. Gitton, P. Anglade, and M. Y. Mistou. 2003. Proteomic analysis of Lactococcus lactis, a lactic acid bacterium. Proteomics 3:337-354.
    • (2003) Proteomics , vol.3 , pp. 337-354
    • Guillot, A.1    Gitton, C.2    Anglade, P.3    Mistou, M.Y.4
  • 12
    • 0037192818 scopus 로고    scopus 로고
    • Structures of nitroreductase in three states: Effects of inhibitor binding and reduction
    • Haynes, C. A., R. L. Koder, A. F. Miller, and D. W. Rodgers. 2002. Structures of nitroreductase in three states: effects of inhibitor binding and reduction. J. Biol. Chem. 277:11513-11520.
    • (2002) J. Biol. Chem. , vol.277 , pp. 11513-11520
    • Haynes, C.A.1    Koder, R.L.2    Miller, A.F.3    Rodgers, D.W.4
  • 13
    • 0031865006 scopus 로고    scopus 로고
    • Touring protein fold space with Dali/FSSP
    • DOI 10.1093/nar/26.1.316
    • Holm, L., and C. Sander. 1998. Touring protein fold space with Dali/FSSP. Nucleic Acids Res. 26:316-319. (Pubitemid 28291549)
    • (1998) Nucleic Acids Research , vol.26 , Issue.1 , pp. 316-319
    • Holm, L.1    Sander, C.2
  • 14
    • 0036015643 scopus 로고    scopus 로고
    • MntR modulates expression of the PerR regulon and superoxide resistance in Staphylococcus aureus through control of manganese uptake
    • Horsburgh, M. J., S. J. Wharton, A. G. Cox, E. Ingham, S. Peacock, and S. J. Foster. 2002. MntR modulates expression of the PerR regulon and superoxide resistance in Staphylococcus aureus through control of manganese uptake. Mol. Microbiol. 44:1269-1286.
    • (2002) Mol. Microbiol. , vol.44 , pp. 1269-1286
    • Horsburgh, M.J.1    Wharton, S.J.2    Cox, A.G.3    Ingham, E.4    Peacock, S.5    Foster, S.J.6
  • 16
    • 17844375077 scopus 로고    scopus 로고
    • Pyruvate protects motor neurons expressing mutant superoxide dismutase 1 against copper toxicity
    • Kim, H. J., J. M. Kim, J. H. Park, J. J. Sung, M. Kim, and K. W. Lee. 2005. Pyruvate protects motor neurons expressing mutant superoxide dismutase 1 against copper toxicity. Neuroreport 16:585-589.
    • (2005) Neuroreport , vol.16 , pp. 585-589
    • Kim, H.J.1    Kim, J.M.2    Park, J.H.3    Sung, J.J.4    Kim, M.5    Lee, K.W.6
  • 17
    • 0035951874 scopus 로고    scopus 로고
    • Structure and site-directed mutagenesis of a flavoprotein from Escherichia coli that reduces nitrocompounds: Alteration of pyridine nucleotide binding by a single amino acid substitution
    • Kobori, T., H. Sasaki, W. C. Lee, S. Zenno, K. Saigo, M. E. Murphy, and M. Tanokura. 2001. Structure and site-directed mutagenesis of a flavoprotein from Escherichia coli that reduces nitrocompounds: alteration of pyridine nucleotide binding by a single amino acid substitution. J. Biol. Chem. 276: 2816-2823.
    • (2001) J. Biol. Chem. , vol.276 , pp. 2816-2823
    • Kobori, T.1    Sasaki, H.2    Lee, W.C.3    Zenno, S.4    Saigo, K.5    Murphy, M.E.6    Tanokura, M.7
  • 18
    • 0015853344 scopus 로고
    • Maturation of the head of bacteriophage T4
    • Laemmli, U. K., and M. Favre. 1973. Maturation of the head of bacteriophage T4. J. Biol. Chem. 80:575-599.
    • (1973) J. Biol. Chem. , vol.80 , pp. 575-599
    • Laemmli, U.K.1    Favre, M.2
  • 19
    • 0032514724 scopus 로고    scopus 로고
    • Mechanism of reduced flavin transfer from Vibrio harveyi NADPH-FMN oxidoreductase to luciferase
    • Lei, B., and S. C. Tu. 1998. Mechanism of reduced flavin transfer from Vibrio harveyi NADPH-FMN oxidoreductase to luciferase. Biochemistry 37:14623-14629.
    • (1998) Biochemistry , vol.37 , pp. 14623-14629
    • Lei, B.1    Tu, S.C.2
  • 20
    • 0029883795 scopus 로고    scopus 로고
    • Copper biochemistry and molecular biology
    • Linder, M. C., and M. Hazegh Azam. 1996. Copper biochemistry and molecular biology. Am. J. Clin. Nutr. 63:797S-811S.
    • (1996) Am. J. Clin. Nutr. , vol.63
    • Linder, M.C.1    Hazegh Azam, M.2
  • 21
    • 0033616612 scopus 로고    scopus 로고
    • Nitroreductase a is regulated as a member of the soxRS regulon of Escherichia coli
    • Liochev, S. I., A. Hausladen, and I. Fridovich. 1999. Nitroreductase A is regulated as a member of the soxRS regulon of Escherichia coli. Proc. Natl. Acad. Sci. U. S. A. 96:3537-3539.
    • (1999) Proc. Natl. Acad. Sci. U. S. A. , vol.96 , pp. 3537-3539
    • Liochev, S.I.1    Hausladen, A.2    Fridovich, I.3
  • 22
    • 66249112833 scopus 로고    scopus 로고
    • The iron-sulfur clusters of dehydratases are primary intracellular targets of copper toxicity
    • Macomber, L., and J. A. Imlay. 2009. The iron-sulfur clusters of dehydratases are primary intracellular targets of copper toxicity. Proc. Natl. Acad. Sci. U. S. A. 106:8344-8349.
    • (2009) Proc. Natl. Acad. Sci. U. S. A. , vol.106 , pp. 8344-8349
    • Macomber, L.1    Imlay, J.A.2
  • 23
    • 38749148524 scopus 로고    scopus 로고
    • Characterization of the CopR regulon of Lactococcus lactis IL1403
    • Magnani, D., O. Barré, S. D. Gerber, and M. Solioz. 2008. Characterization of the CopR regulon of Lactococcus lactis IL1403. J. Bacteriol. 190:536-545.
    • (2008) J. Bacteriol. , vol.190 , pp. 536-545
    • Magnani, D.1    Barré, O.2    Gerber, S.D.3    Solioz, M.4
  • 24
    • 0033986006 scopus 로고    scopus 로고
    • Increased production of hydrogen peroxide by Lactobacillus delbrueckii subsp. bulgaricus upon aeration: Involvement of an NADH oxidase in oxidative stress
    • Marty-Teysset, C., F. de la Torre, and J. Garel. 2000. Increased production of hydrogen peroxide by Lactobacillus delbrueckii subsp. bulgaricus upon aeration: involvement of an NADH oxidase in oxidative stress. Appl. Environ. Microbiol. 66:262-267.
    • (2000) Appl. Environ. Microbiol. , vol.66 , pp. 262-267
    • Marty-Teysset, C.1    De La Torre, F.2    Garel, J.3
  • 26
    • 0031694118 scopus 로고    scopus 로고
    • Virus directed enzyme prodrug therapy for ovarian and pancreatic cancer using retrovirally delivered E. coli nitroreductase and CB1954
    • McNeish, I. A., N. K. Green, M. G. Gilligan, M. J. Ford, V. Mautner, L. S. Young, D. J. Kerr, and P. F. Searle. 1998. Virus directed enzyme prodrug therapy for ovarian and pancreatic cancer using retrovirally delivered E. coli nitroreductase and CB1954. Gene Ther. 5:1061-1069.
    • (1998) Gene Ther. , vol.5 , pp. 1061-1069
    • McNeish, I.A.1    Green, N.K.2    Gilligan, M.G.3    Ford, M.J.4    Mautner, V.5    Young, L.S.6    Kerr, D.J.7    Searle, P.F.8
  • 28
    • 1242318677 scopus 로고    scopus 로고
    • Transcriptome and proteome analysis of Bacillus subtilis gene expression in response to superoxide and peroxide stress
    • Mostertz, J., C. Scharf, M. Hecker, and G. Homuth. 2004. Transcriptome and proteome analysis of Bacillus subtilis gene expression in response to superoxide and peroxide stress. Microbiology 150:497-512.
    • (2004) Microbiology , vol.150 , pp. 497-512
    • Mostertz, J.1    Scharf, C.2    Hecker, M.3    Homuth, G.4
  • 29
    • 1642514907 scopus 로고    scopus 로고
    • Prominent roles of the NorR and fur regulators in the Escherichia coli transcriptional response to reactive nitrogen species
    • Mukhopadhyay, P., M. Zheng, L. A. Bedzyk, R. A. LaRossa, and G. Storz. 2004. Prominent roles of the NorR and Fur regulators in the Escherichia coli transcriptional response to reactive nitrogen species. Proc. Natl. Acad. Sci. U. S. A. 101:745-750.
    • (2004) Proc. Natl. Acad. Sci. U. S. A. , vol.101 , pp. 745-750
    • Mukhopadhyay, P.1    Zheng, M.2    Bedzyk, L.A.3    LaRossa, R.A.4    Storz, G.5
  • 30
    • 0028938093 scopus 로고
    • Two trans-acting metalloregulatory proteins controlling expression of the copper-ATPases of Enterococcus hirae
    • Odermatt, A., and M. Solioz. 1995. Two trans-acting metalloregulatory proteins controlling expression of the copper-ATPases of Enterococcus hirae. J. Biol. Chem. 270:4349-4354.
    • (1995) J. Biol. Chem. , vol.270 , pp. 4349-4354
    • Odermatt, A.1    Solioz, M.2
  • 33
    • 0023472472 scopus 로고
    • Tricine-sodium dodecyl sulfate-olyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa
    • Schagger, H., and G. von Jagow. 1987. Tricine-sodium dodecyl sulfate-olyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal. Biochem. 166:368-379.
    • (1987) Anal. Biochem. , vol.166 , pp. 368-379
    • Schagger, H.1    Von Jagow, G.2
  • 34
    • 0037565100 scopus 로고    scopus 로고
    • Copper homeostasis in Enterococcus hirae
    • Solioz, M., and J. V. Stoyanov. 2003. Copper homeostasis in Enterococcus hirae. FEMS Microbiol. Rev. 27:183-195.
    • (2003) FEMS Microbiol. Rev. , vol.27 , pp. 183-195
    • Solioz, M.1    Stoyanov, J.V.2
  • 35
    • 0025012990 scopus 로고
    • Manganese-dependent disproportionation of hydrogen peroxide in bicarbonate buffer
    • Stadtman, E. R., B. S. Berlett, and P. B. Chock. 1990. Manganese-dependent disproportionation of hydrogen peroxide in bicarbonate buffer. Proc. Natl. Acad. Sci. U. S. A. 87:384-388.
    • (1990) Proc. Natl. Acad. Sci. U. S. A. , vol.87 , pp. 384-388
    • Stadtman, E.R.1    Berlett, B.S.2    Chock, P.B.3
  • 36
    • 0031002899 scopus 로고    scopus 로고
    • CopY is a copper-inducible repressor of the Enterococcus hirae copper ATPases
    • Strausak, D., and M. Solioz. 1997. CopY is a copper-inducible repressor of the Enterococcus hirae copper ATPases. J. Biol. Chem. 272:8932-8936.
    • (1997) J. Biol. Chem. , vol.272 , pp. 8932-8936
    • Strausak, D.1    Solioz, M.2
  • 37
    • 15244355729 scopus 로고    scopus 로고
    • Staphylococcus aureus NfrA (SA0367) is a flavin mononucleotide-dependent NADPH oxidase involved in oxidative stress response
    • Streker, K., C. Freiberg, H. Labischinski, J. Hacker, and K. Ohlsen. 2005. Staphylococcus aureus NfrA (SA0367) is a flavin mononucleotide-dependent NADPH oxidase involved in oxidative stress response. J. Bacteriol. 187: 2249-2256.
    • (2005) J. Bacteriol. , vol.187 , pp. 2249-2256
    • Streker, K.1    Freiberg, C.2    Labischinski, H.3    Hacker, J.4    Ohlsen, K.5
  • 38
    • 33846946121 scopus 로고    scopus 로고
    • Synechocystis DrgA protein functioning as nitroreductase and ferric reductase is capable of catalyzing the Fenton reaction
    • Takeda, K., M. Iizuka, T. Watanabe, J. Nakagawa, S. Kawasaki, and Y. Niimura. 2007. Synechocystis DrgA protein functioning as nitroreductase and ferric reductase is capable of catalyzing the Fenton reaction. FEBS J. 274: 1318-1327.
    • (2007) FEBS J. , vol.274 , pp. 1318-1327
    • Takeda, K.1    Iizuka, M.2    Watanabe, T.3    Nakagawa, J.4    Kawasaki, S.5    Niimura, Y.6
  • 39
    • 0016686551 scopus 로고
    • Improved medium for lactic streptococci and their bacteriophages
    • Terzaghi, B. E., and W. E. Sandine. 1975. Improved medium for lactic streptococci and their bacteriophages. Appl. Microbiol. 29:807-813.
    • (1975) Appl. Microbiol. , vol.29 , pp. 807-813
    • Terzaghi, B.E.1    Sandine, W.E.2
  • 40
    • 33751518049 scopus 로고    scopus 로고
    • Composition and catalase-like activity of Mn(II)-bicarbonate complexes
    • Mosc.
    • Tikhonov, K. G., O. M. Zastrizhnaya, Y. N. Kozlov, and V. V. Klimov. 2006. Composition and catalase-like activity of Mn(II)-bicarbonate complexes. Biochemistry (Mosc.) 71:1270-1277.
    • (2006) Biochemistry , vol.71 , pp. 1270-1277
    • Tikhonov, K.G.1    Zastrizhnaya, O.M.2    Kozlov, Y.N.3    Klimov, V.V.4
  • 41
    • 0009482260 scopus 로고
    • Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications
    • Towbin, H., T. Staehelin, and J. Gordon. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. U. S. A. 76:4350-4354.
    • (1979) Proc. Natl. Acad. Sci. U. S. A. , vol.76 , pp. 4350-4354
    • Towbin, H.1    Staehelin, T.2    Gordon, J.3
  • 43
    • 24044481966 scopus 로고    scopus 로고
    • Copper, oxidative stress, and human health
    • Uriu-Adams, J. Y., and C. L. Keen. 2005. Copper, oxidative stress, and human health. Mol. Aspects Med. 26:268-298.
    • (2005) Mol. Aspects Med. , vol.26 , pp. 268-298
    • Uriu-Adams, J.Y.1    Keen, C.L.2
  • 45
    • 0036729482 scopus 로고    scopus 로고
    • Formation and conversion of oxygen metabolites by Lactococcus lactis subsp. lactis ATCC 19435 under different growth conditions
    • van Niel, E. W., K. Hofvendahl, and B. Hahn-Hagerdal. 2002. Formation and conversion of oxygen metabolites by Lactococcus lactis subsp. lactis ATCC 19435 under different growth conditions. Appl. Environ. Microbiol. 68:4350-4356.
    • (2002) Appl. Environ. Microbiol. , vol.68 , pp. 4350-4356
    • Van Niel, E.W.1    Hofvendahl, K.2    Hahn-Hagerdal, B.3
  • 46
    • 0035873398 scopus 로고    scopus 로고
    • Pyruvate released by astrocytes protects neurons from copper-catalyzed cysteine neurotoxicity
    • Wang, X. F., and M. S. Cynader. 2001. Pyruvate released by astrocytes protects neurons from copper-catalyzed cysteine neurotoxicity. J. Neurosci. 21:3322-3331.
    • (2001) J. Neurosci. , vol.21 , pp. 3322-3331
    • Wang, X.F.1    Cynader, M.S.2
  • 47
    • 33750348724 scopus 로고    scopus 로고
    • Characterization of genes involved in the initial reactions of 4-chloronitrobenzene degradation in Pseudomonas putida ZWL73
    • Xiao, Y., J. F. Wu, H. Liu, S. J. Wang, S. J. Liu, and N. Y. Zhou. 2006. Characterization of genes involved in the initial reactions of 4-chloronitrobenzene degradation in Pseudomonas putida ZWL73. Appl. Microbiol. Biotechnol. 73:166-171.
    • (2006) Appl. Microbiol. Biotechnol. , vol.73 , pp. 166-171
    • Xiao, Y.1    Wu, J.F.2    Liu, H.3    Wang, S.J.4    Liu, S.J.5    Zhou, N.Y.6
  • 48
    • 0029745672 scopus 로고    scopus 로고
    • Biochemical characterization of NfsA, the Escherichia coli major nitroreductase exhibiting a high amino acid sequence homology to Frp, a Vibrio harveyi flavin oxidoreductase
    • Zenno, S., H. Koike, A. N. Kumar, R. Jayaraman, M. Tanokura, and K. Saigo. 1996. Biochemical characterization of NfsA, the Escherichia coli major nitroreductase exhibiting a high amino acid sequence homology to Frp, a Vibrio harveyi flavin oxidoreductase. J. Bacteriol. 178:4508-4514.
    • (1996) J. Bacteriol. , vol.178 , pp. 4508-4514
    • Zenno, S.1    Koike, H.2    Kumar, A.N.3    Jayaraman, R.4    Tanokura, M.5    Saigo, K.6
  • 49
    • 0028206223 scopus 로고
    • Identification of the genes encoding NAD(P)H-flavin oxidoreductases that are similar in sequence to Escherichia coli Fre in four species of luminous bacteria: Photorhabdus luminescens, Vibrio fischeri, Vibrio harveyi, and Vibrio orientalis
    • Zenno, S., and K. Saigo. 1994. Identification of the genes encoding NAD(P)H-flavin oxidoreductases that are similar in sequence to Escherichia coli Fre in four species of luminous bacteria: Photorhabdus luminescens, Vibrio fischeri, Vibrio harveyi, and Vibrio orientalis. J. Bacteriol. 176:3544-3551.
    • (1994) J. Bacteriol. , vol.176 , pp. 3544-3551
    • Zenno, S.1    Saigo, K.2


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.