SCOPUS 정보 검색 플랫폼

Progress in Nuclear Magnetic Resonance Spectroscopy

Volumn 39, Issue 3, 2001, Pages 171-190

Labelling approaches for protein structural studies by solution-state and solid-state NMR

(2) Lian, L Y a Middleton, D A a

a UNIVERSITY OF MANCHESTER (United Kingdom)

Author keywords

Deuteration; Isotope labelling; NMR; Protein expression; Proteins; Spin labelling; Splicing

Indexed keywords

ISOTOPES; MACROMOLECULES; MOLECULAR STRUCTURE; NUCLEAR MAGNETIC RESONANCE; PROTEINS;

ISOTOPE LABELING; SPIN LABELING;

SOLID STATE PHYSICS;

EID: 0035913884 PISSN: 00796565 EISSN: None Source Type: Journal
DOI: 10.1016/S0079-6565(01)00034-6 Document Type: Review

Times cited : (106)

References (115)

1
- 0030749281
- (1997) Nat. Struct. Biol. Suppl. , vol.4 , pp. 858-861
- Kainosho, M.¹

2
- 0033794450
- (2000) Curr. Opin. Struct. Biol. , vol.10 , pp. 585-592
- Goto, N.K.¹ Kay, L.E.²

3
- 0032486760
- (1998) J. Am. Chem. Soc. , vol.120 , pp. 7617-7625
- Zwahlen, C.¹ Gardner, K.H.² Sarma, S.P.³ Horita, D.A.⁴ Byrd, R.A.⁵ Kay, L.E.⁶

4
- 0000187904
- (1996) J. Biomol. NMR , vol.8 , pp. 360-368
- Smith, B.O.¹ Ito, Y.² Raine, A.³ Teichmann, S.⁴ Ben-Tovim, L.⁵ Nietlispach, D.⁶ Broadshurst, R.W.⁷ Terada, T.⁸ Kelly, M.⁹ Oschkinat, H.¹⁰ Shibata, T.¹¹ Yokoyama, S.¹² Laue, E.D.¹³

5
- 0029921643
- (1996) Biochemistry , vol.35 , pp. 6116-6125
- Wand, A.J.¹ Urbauer, J.L.² McEvoy, R.P.³ Bieber, R.J.⁴

6
- 0029905210
- (1996) J. Am. Chem. Soc. , vol.118 , pp. 9255-9264
- LeMaster, D.M.¹ Kushlan, D.M.²

7
- 0029176895
- (1995) J. Am. Chem. Soc. , vol.117 , pp. 11536-11544
- Muhandiram, D.R.¹ Yamazaki, T.² Sykes, B.D.³ Kay, L.E.⁴

8
- 0030921081
- (1997) J. Am. Chem. Soc. , vol.119 , pp. 3842-3843
- Pervushin, K.¹ Wider, G.² Wuthrich, K.³

9
- 0033572874
- (1999) J. Am. Chem. Soc. , vol.121 , pp. 11589-11590
- Ishima, R.¹ Loius, J.M.² Torchia, D.A.³

10
- 0033497114
- (2000) Clin. Chem. Enzymol. Commun. J. , vol.8 , pp. 239-253
- Allardyce, C.S.¹ McDonagh, P.D.² Prescott, A.³ Roberts, G.C.K.⁴ Lian, L.Y.⁵

11
- 0030612833
- (1997) Proc. Natl Acad. Sci. USA , vol.94 , pp. 12366-12371
- Pervushin, K.¹ Riek, R.² Wider, G.³ Wuthrich, K.⁴

12
- 0031595590
- (1998) Ann. Rev. Biophys. Biomol. Struct. , vol.27 , pp. 357-406
- Gardner, K.H.¹ Kay, L.E.²

13
- 0346672683
- (2000) Phil. Trans. R. Soc. Lond. A , vol.358 , pp. 513-545
- Pfuhl, M.¹ Driscoll, P.C.²

14
- 0030589110
- (1996) Curr. Biol. , vol.4 , pp. 1245-1249
- Sattler, M.¹ Fesik, S.W.²

15
- 0001388945
- (1998) Prog. NMR Spectrosc. , vol.32 , pp. 277-286
- Arrowsmith, C.H.¹ Wu, Y.S.²

16
- 0033121307
- (1999) J. Biomol. NMR , vol.14 , pp. 71-74
- Hong, M.¹ Jakes, K.²

17
- 0033998371
- (2000) J. Biomol. NMR , vol.16 , pp. 209-219
- McDermott, A.¹ Polenova, T.² Bockmann, A.³ Zilm, K.⁴ Paulsen, E.K.⁵ Martin, R.W.⁶ Montelione, G.T.⁷

18
- 0034167578
- (2000) J. Magn. Reson. , vol.143 , pp. 411-416
- Pauli, J.¹ Van Rossum, B.² Forster, H.³ De Groot, H.J.⁴ Oschkinat, H.⁵

19
- 0031764019
- (1998) Curr. Opin. Struct. Biol. , vol.8 , pp. 640-648
- Marassi, F.M.¹ Opella, S.J.²

20
- 0030844584
- (1997) Proc. Natl Acad. Sci. USA , vol.94 , pp. 8551-8556
- Marassi, F.M.¹ Ramamoorthy, A.² Opella, S.J.³

21
- 67049170125
- PhD Thesis. University of Leicester, UK
- (2000)
- Verdemato, P.E.¹

22
- 0031627465
- (1998) J. Biomol. NMR , vol.11 , pp. 97-102
- Cai, M.¹ Huang, Y.² Sakaguchi, K.³ Clore, G.M.⁴ Gronenborn, A.M.⁵ Craigie, R.⁶

23
- 0034307588
- (2000) Nucl. Acids Res. , vol.28 , pp. 3710-3718
- Verdemato, P.E.¹ Brannigan, J.A.² Damblon, C.F.³ Moody, P.C.E.⁴ Zuccotto, F.⁵ Lian, L.Y.⁶

24
- 67049166494
- ASM
- (1996)
- Neidhardt, F.C.¹ Curtiss, R.² Ingraham, J.L.³ Liu, E.C.C.⁴ Low, K.B.⁵ Magasanik, B.⁶ Reznikoff, W.G.⁷ Riley, M.⁸ Schaechter, M.⁹ Umbarger, H.E.¹⁰

25
- 0031554635
- (1997) Biotechnol. Bioengng , vol.55 , pp. 101-117
- Wiechert, W.¹ DeGraaf, A.A.²

26
- 0031554628
- (1997) Biotechnol. Bioengng , vol.55 , pp. 118-1134
- Wiechert, W.¹ Siefke, C.² DeGraaf, A.A.³ Marx, A.⁴

27
- 0033155962
- (1999) Metabol. Engng , vol.1 , pp. 189-197
- Szyperski, T.¹ Glaser, R.W.² Hochuli, M.³ Fiaux, J.⁴

28
- 0033524958
- (1999) J. Mol. Biol. , vol.285 , pp. 2119-2132
- McCallum, S.A.¹ Hitchens, T.K.² Rule, G.S.³

29
- 0032753923
- (1999) J. Bacteriol. , vol.181 , pp. 6679-6688
- Sauer, U.¹ Lasko, D.R.² Fiau, J.³ Hochuli, M.⁴ Glaser, R.⁵ Szyperski, T.⁶ Wuthrich, K.⁷ Bailey, J.E.⁸

30
- 0034706029
- (2000) J. Am. Chem. Soc. , vol.122 , pp. 7898-7904
- Hajduk, P.J.¹ Augeri, D.J.² Mack, J.³ Mendoza, R.⁴ Yang, J.⁵ Betz, S.F.⁶ Fesik, S.W.⁷

31
- 0030824756
- (1997) J. Am. Chem. Soc. , vol.119 , pp. 7599-7600
- Gardner, K.H.¹ Kay, L.E.²

32
- 0032898682
- (1999) J. Biomol. NMR , vol.13 , pp. 369-374
- Goto, N.K.¹ Gardner, K.H.² Mueller, G.A.³ Willis, R.C.⁴ Kay, L.E.⁵

33
- 0031154310
- (1997) J. Biomol. NMR , vol.9 , pp. 437-440
- Lee, A.L.¹ Urbauer, J.L.² Wand, A.J.³

34
- 0033170449
- (1999) J. Magn. Reson. , vol.139 , pp. 389-401
- Hong, M.¹

35
- 0030589148
- (1996) J. Mol. Biol. , vol.263 , pp. 627-636
- Rosen, M.K.¹ Gardner, K.H.² Willis, R.³ Parris, W.E.⁴ Pawson, T.⁵ Kay, L.E.⁶

36
- 0004049181
- (1992) Methods Enzymol. , pp. 44-73
- Muchmore, D.C.¹ McIntosh, L.P.² Russell, C.B.³ Anderson, D.E.⁴ Dahlquist, F.W.⁵

37
- 0030239071
- (1996) J. Biomol. NMR , vol.8 , pp. 184-192
- Waugh, D.S.¹

38
- 0030939030
- (1997) Biochemistry , vol.36 , pp. 3404-3416
- Duewel, H.¹ Daub, E.² Robinson, V.³ Honek, J.F.⁴

39
- 0032485881
- (1998) Biochemistry , vol.37 , pp. 6286-6294
- Parsons, J.F.¹ Xiao, G.² Gilliland, G.L.³ Armstrong, R.N.⁴

40
- 0031985317
- (1998) Nat. Struct. Biol. , vol.5 , pp. 3-5
- Stoddard, B.L.¹ Pietrokovski, S.²

41
- 0031975772
- (1998) Nat. Struct. Biol. , vol.5 , pp. 31-36
- Klabunde, T.¹ Sharma, S.² Telenti, A.³ Jacobs, W.R.⁴ Sacchettini, J.C.⁵

42
- 0030611387
- (1997) Cell , vol.89 , pp. 555-564
- Duan, X.¹ Gimble, F.S.² Quiocho, F.A.³

43
- 0032193937
- (1998) Chem. Biol. , vol.5
- Gimble, F.S.¹

44
- 0032503612
- (1998) J. Am. Chem. Soc. , vol.120 , pp. 5591-5592
- Yamazaki, T.¹ Otomo, T.² Oda, N.³ Kyogoku, Y.⁴ Uegaki, K.⁵ Ito, N.⁶ Ishino, Y.⁷ Nakamura, H.⁸

45
- 0033038793
- (1999) J. Biomol. NMR , vol.14 , pp. 105-114
- Otomo, T.¹ Teruya, K.² Uegaki, K.³ Yamazaki, T.⁴ Kyogoku, Y.⁵

46
- 0033534179
- (1999) Biochemistry , vol.38 , pp. 16040-16044
- Otomo, T.¹ Ito, N.² Kyogoku, Y.³ Yamazaki, T.⁴

47
- 0033582287
- (1999) Proc. Natl Acad. Sci. USA , vol.96 , pp. 388-393
- Xu, R.¹ Ayers, B.² Cowburn, D.³ Muir, T.W.⁴

48
- 0032499752
- (1998) Proc. Natl Acad. Sci. USA , vol.95 , pp. 6705-6710
- Muir, T.W.¹ Sondhi, D.² Cole, P.A.³

49
- 0032568924
- (1998) J. Biol. Chem. , vol.273 , pp. 16205-16209
- Severinov, K.¹ Muir, T.W.²

50
- 0344351815
- (1998) Protein Sci. , vol.7 , pp. 2256-2264
- Evans T.C., Jr.¹ Brenner, J.² Xu, M.Q.³

51
- 0032133387
- (1998) Biomol. NMR , vol.12 , pp. 259-276
- Mal, T.K.¹ Matthews, S.J.² Kovacs, H.³ Campbell, I.D.⁴ Boyd, J.J.⁵

52
- 0030902818
- (1997) Nat. Struct. Biol. , vol.4 , pp. 483-489
- Yu, L.¹ Petros, A.M.² Schnuchel, A.³ Zhong, P.⁴ Severin, J.M.⁵ Walter, K.⁶ Holzman, T.F.⁷ Fesik, S.W.⁸

53
- 0028520420
- (1994) J. Magn. Reson. Ser. B , vol.105 , pp. 192-195
- Markus, M.A.¹ Dayie, K.T.² Matsudaira, P.³ Wagner, G.⁴

54
- 0030596171
- (1996) J. Mol. Biol. , vol.264 , pp. 1101-1116
- Venters, R.A.¹ Farmer, B.T.² Fierke, C.A.³ Spicer, L.D.⁴

55
- 0033136436
- (1999) J. Biomol. NMR , vol.14 , pp. 79-83
- Kelly, M.J.S.¹ Krieger, C.² Ball, L.J.³ Yu, Y.⁴ Richter, G.⁵ Schmieder, P.⁶ Bacher, A.⁷ Oschkinat, H.⁸

56
- 0029317862
- (1995) J. Biomol. NMR , vol.5 , pp. 339-344
- Venters, R.A.¹ Huang, C.C.² Farmer, B.T.³ Trolard, R.⁴ Spicer, L.D.⁵ Fierke, C.A.⁶

57
- 0001258823
- (1994) Prog. Nucl. Magn. Reson. Spectrosc. , vol.26 , pp. 371-419
- LeMaster, D.M.¹

58
- 0032534788
- (1998) Anal. Biochem. , vol.265 , pp. 351-355
- Leiting, B.¹ Marsilio, F.² O'Connell, J.F.³

59
- 13344295061
- (1996) J. Am. Chem. Soc. , vol.118 , pp. 407-415
- Nietlispach, D.¹ Clowes, R.T.² Broadhurst, R.W.³ Ito, Y.⁴ Keeler, J.⁵ Kelly, M.⁶ Ashurst, J.⁷ Oschkinat, H.⁸ Domaille, P.J.⁹ Laue, E.¹⁰

60
- 0033918093
- (2000) J. Biomol. NMR , vol.17 , pp. 33-42
- Hochuli, M.¹ Szyperski, T.² Wuthrich, K.³

61
- 67049123854
- unpublished work
- (1998)
- Lian, L.Y.¹ Prescott, A.² Roberts, G.C.K.³ Cullis, P.⁴

62
- 0026536266
- (1992) Biotechnol. Lett. , vol.14 , pp. 269-274
- Chen, S.T.¹ Tu, C.C.² Wang, K.T.³

63
- 0027994518
- (1994) J. Label. Compd Radiopharm. , vol.34 , pp. 831-834
- Nishiyama, K.¹ Oba, M.² Ueno, R.³ Morita, A.⁴ Nakamura, Y.⁵ Kainosho, M.⁶

64
- 0029997736
- (1996) J. Am. Chem. Soc. , vol.118 , pp. 6800-6801
- Metzler, W.J.¹ Wittekind, M.² Goldfarb, V.³ Mueller, L.⁴ Farmer, B.T.⁵

65
- 0028934760
- (1995) J. Amino Acids , vol.8 , pp. 171-186
- Raap, J.¹ Wolthuis, W.N.E.² Hehenkamp, J.J.J.³ Lugtenburg, J.⁴

66
- 0028858278
- (1995) J. Label. Compd Radiopharm. , vol.36 , pp. 1077-1096
- Karstens, W.F.J.¹ Berger, H.J.F.F.² Van Haren, E.R.³ Lugtenburg, J.⁴ Raap, J.⁵

67
- 0000816468
- (1994) Recl. Trav. Chim. Pays-Bas-J. R. Neth. Chem. Soc. , vol.113 , pp. 369-375
- Ogrel, A.¹ Vasilanko, I.A.² Lugtenburg, J.³ Raap, J.⁴

68
- 0028930179
- (1995) J. Label. Compd Radiopharm. , vol.36 , pp. 439-444
- Panigot, M.J.¹ Fesik, S.W.² Curley, R.W.³ Raap, J.⁴

69
- 0031937870
- (1998) Protein Sci. , vol.7 , pp. 419-426
- Furter, R.¹

70
- 0028483441
- (1994) Nat. Struct. Biol. , vol.1 , pp. 512-517
- Sonar, S.¹ Lee, C.P.² Coleman, M.³ Patel, N.⁴ Liu, X.M.⁵ Marti, T.⁶ Khorana, H.G.⁷ Rajbhandary, U.L.⁸ Rothschild, K.J.⁹

71
- 0032036838
- (1998) J. Biomol. NMR , vol.11 , pp. 295-306
- Yabuki, T.¹ Kigawa, T.² Dohmae, N.³ Takio, K.⁴ Terada, T.⁵ Ito, Y.⁶ Laue, E.D.⁷ Cooper, J.A.⁸ Kainosho, M.⁹ Yokoyama, S.¹⁰

72
- 0029142564
- (1995) Rev. Biophys. , vol.28 , pp. 315-422
- Grisshammer, R.¹ Tate, C.G.Q.²

73
- 0030437935
- (1996) Q. Rev. Biophys. , vol.29 , pp. 395-449
- Smith, S.O.¹ Aschheim, K.² Groesbeek, M.Q.³

74
- 0033776959
- (2000) Curr. Opin. Chem. Biol. , vol.4 , pp. 500-506
- Tycko, R.¹

75
- 0034073419
- (2000) J. Biomol. NMR , vol.16 , pp. 313-327
- Balbach, J.J.¹ Yang, J.² Weliky, D.P.³ Steinbach, P.J.⁴ Tugarinov, V.⁵ Anglister, J.⁶ Tycko, R.⁷

76
- 0003860475
- Pierce Chemical Company Rockford, IL
- (1984) Solid Phase Peptide Synthesis, 2nd ed
- Stewart, J.M.¹ Young, J.D.²

77
- 0032704310
- (1999) J. Pept. Res. , vol.54 , pp. 427-453
- Cornille, F.¹ Wecker, K.² Loffet, A.³ Genet, R.⁴ Roques, B.⁵

78
- 0030696099
- (1997) Meth. Enzymol. , vol.289 , pp. 697-717
- Angelleti, R.H.¹ Boewald, L.F.² Fields, G.B.³

79
- 0030711495
- (1997) Meth. Enzymol. , vol.289 , pp. 266-298
- Muir, T.W.¹ Dawson, P.E.² Kent, S.B.³

80
- 0032757152
- (1999) Biopolymers , vol.51 , pp. 174-190
- Bechinger, B.¹ Kinder, R.² Helmle, M.³ Vogt, T.C.⁴ Harzer, U.⁵ Schinzel, S.⁶

81
- 44949291859
- (1990) J. Magn. Reson. , vol.90 , pp. 330-340
- Pan, Y.¹ Gullion, T.² Schaefer, J.³

82
- 36549091040
- (1990) J. Chem. Phys. , vol.92 , pp. 6347-6364
- Levitt, M.H.¹ Raleigh, D.P.² Creuzet, F.³ Griffin, R.G.⁴

83
- 0000528084
- (1995) J. Am. Chem. Soc. , vol.117 , pp. 7228-7237
- Peersen, O.B.¹ Groesbeek, M.² Aimoto, S.³ Smith, S.O.⁴

84
- 67049111793
- DPhil Thesis. University of Oxford
- (2000)
- Ahmed, Z.¹

85
- 0027219740
- (1993) Biochemistry , vol.32 , pp. 7593-7604
- Hing, A.W.¹ Schaefer, J.²

86
- 0029794277
- (1996) Biochemistry , vol.35 , pp. 12733-12741
- Hirsh, D.J.¹ Hammer, J.² Maloy, W.L.³ Blazyk, J.⁴ Schaefer, J.⁵

87
- 0034220429
- (2000) J. Magn. Reson. , vol.145 , pp. 132-141
- Reif, B.¹ Hohwy, M.² Jaroniec, C.P.³ Reinstra, C.⁴ Griffin, R.G.⁵

88
- 0034730691
- (2000) Biochim. Biophys. Acta , vol.1468 , pp. 187-198
- Ahmed, Z.¹ Watts, A.² Middleton, D.A.³

89
- 0343185890
- (2000) Biochim. Biophys. Acta , vol.1463 , pp. 151-161
- Glaubitz, C.¹ Gröbner, G.² Watts, A.³

90
- 0034000232
- (2000) J. Biomol. NMR , vol.16 , pp. 261-268
- Fu, R.¹ Cotton, M.² Cross, T.A.³

91
- 0028814024
- (1995) Proc. Natl Acad. Sci. USA , vol.92 , pp. 488-491
- Smith, S.O.¹ Bormann, B.J.²

92
- 0034700129
- (2000) Proc. Natl Acad. Sci. USA , vol.97 , pp. 13045-13050
- Antzutkin, O.N.¹ Balbach, J.J.² Leapman, R.D.³ Rizzo, N.W.⁴ Reed, J.⁵ Tycko, R.⁶

93
- 0028804827
- (1995) Nat. Struct. Biol. , vol.2 , pp. 990-998
- Lansbury, P.T.¹ Costa, P.R.² Griffiths, J.M.³ Simon, E.J.⁴ Auger, M.⁵ Halverson, K.J.⁶ Kocisko, D.A.⁷ Hendsch, Z.S.⁸ Ashburn, T.T.⁹ Spencer, R.G.S.¹⁰ Tidor, B.¹¹ Griffin, R.G.¹²

94
- 67049170124
- CRC Press, Boca Raton
- (1995)
- Lundblad, R.L.¹

95
- 0024853292
- (1989) Meth. Enzymol. , vol.177 , pp. 86-121
- Kosen, P.A.¹

96
- 0020669729
- (1983) Meth. Enzymol. , vol.19 , pp. 570-579
- Jentoft, N.¹ Dearborn, D.P.G.²

97
- 0025910862
- (1991) Biochemistry , vol.30 , pp. 3871-3879
- Spooner, P.J.¹ Watts, A.²

98
- 0029332135
- (1995) Mol. Membr. Biol. , vol.12 , pp. 233-246
- Watts, A.¹ Ulrich, A.S.² Middleton, D.A.³

99
- 0022559063
- (1986) Biochemistry , vol.25 , pp. 2187-2191
- Brauer, M.¹ Sykes, B.D.²

100
- 0028500413
- (1994) Enzyme Microb. Technol. , vol.16 , pp. 795-801
- Richards, P.G.¹ Coles, B.² Heptinstall, J.³ Walton, D.J.⁴

101
- 0033812585
- (2000) Nat. Struct. Biol. , vol.7 , pp. 735-739
- Hubbel, W.L.¹ Cafiso, D.S.² Altenbach, C.³

102
- 0034705338
- (2000) J. Mol. Biol. , vol.299 , pp. 1341-1351
- Yi, Q.¹ Scalley-Kim, M.L.² Alm, E.J.³ Baker, D.⁴

103
- 0034625121
- (2000) Biochemistry , vol.39 , pp. 5355-5365
- Battiste, J.L.¹ Wagner, G.²

104
- 0034695426
- (2000) J. Mol. Biol. , vol.295 , pp. 595-603
- Garcia, F.L.¹ Szyperski, T.² Dyer, J.H.³ Choinowski, T.⁴ Seedorf, U.⁵ Hauser, H.⁶ Wüthrich, K.⁷

105
- 0033609437
- (1999) Biochemistry , vol.38 , pp. 9634-9639
- Spooner, P.J.¹ Veenhoff, L.M.² Watts, A.³ Poolman, B.⁴

106
- 0033515440
- (1999) J. Mol. Biol. , vol.287 , pp. 609-625
- Johnson, P.E.¹ Brun, E.² MacKenzie, L.F.³ Withers, S.G.⁴ McIntosh, L.P.⁵

107
- 0032042825
- (1998) J. Magn. Reson. , vol.131 , pp. 248-253
- Bolin, K.A.¹ Hanson, P.² Wright, S.J.³ Millhauser, G.L.⁴

108
- 0001583001
- (1995) Biochemistry , vol.34 , pp. 1645-1653
- Girvin, M.E.¹ Fillingame, R.H.²

109
- 0031585990
- (1997) J. Mol. Biol. , vol.268 , pp. 158-169
- Gillespie, J.R.¹ Shortie, D.²

110
- 36849133491
- (1956) J. Chem. Phys. , vol.25 , pp. 261-266
- Solomon, I.¹ Bloembergen, N.²

111
- 0028801713
- (1995) FASEB J. , vol.9 , pp. 120-126
- Scherf, T.¹ Hiller, R.² Anglister, J.³

112
- 0034822661
- (2001) J. Am. Chem. Soc. , vol.123 , pp. 3149-3150
- Jahnke, W.¹ Rüisser, S.² Zurini, M.³

113
- 0025923891
- (1991) Biochemistry , vol.30 , pp. 4491-4494
- Venters, R.A.¹ Calderone, T.L.² Spicer, L.D.³ Fierke, C.A.⁴

114
- 0032134275
- (1998) J. Biomol. NMR , vol.12 , pp. 299-306
- Jasanoff, A.¹

115
- 0026705562
- (1992) Biochemistry , vol.31 , pp. 8191-8195
- Derrick, J.P.¹ Lian, L.Y.² Roberts, G.C.K.³ Shaw, W.V.⁴

* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.