Microsecond timescale backbone conformational dynamics in ubiquitin studied with NMR R1rho relaxation experiments.
NMR spin relaxation experiments are used to characterize the dynamics of the backbone of ubiquitin. Chemical exchange processes affecting residues Ile 23, Asn 25, Thr 55, and Val 70 are characterized using on- and off-resonance rotating-frame 15N R1rho relaxation experiments to have a kinetic exchange rate constant of 25,000 sec(-1) at 280 K. The exchange process affecting residues 23, 25, and 55 appears to result from disruption of N-cap hydrogen bonds of the alpha-helix and possibly from repacking of the side chain of Ile 23. Chemical exchange processes affecting other residues on the surface of ubiquitin are identified using 1H-15N multiple quantum relaxation experiments. These residues are located near or at the regions known to interact with various enzymes of the ubiquitin-dependent protein degradation pathway.
Massi
Francesca
F
Department of Biochemistry and Molecular Biophysics, Columbia University, 630 West 168th Street, New York, NY 10032, USA.
Grey
Michael J
MJ
Palmer
Arthur G
AG
D009682:Magnetic Resonance Spectroscopy; D009587:Nitrogen Isotopes; D017434:Protein Structure, Tertiary; D025801:Ubiquitin / Q000737:chemistry*
D009587:Nitrogen Isotopes; D025801:Ubiquitin
Biochemistry. 1994 May 17;33(19):5984-6003
7514039
J Biomol NMR. 2000 Dec;18(4):319-27
11200526
Annu Rev Biochem. 1998;67:425-79
9759494
J Mol Biol. 2002 Apr 12;317(5):739-52
11955021
Biochemistry. 1994 Jun 14;33(23):7300-8
8003494
J Am Chem Soc. 2003 Nov 26;125(47):14324-35
14624581
J Biomol NMR. 1995 Nov;6(3):277-93
8520220
J Biomol NMR. 2003 Mar;25(3):173-95
12652131
J Biomol NMR. 2004 Mar;28(3):213-27
14752255
J Biomol NMR. 2002 May;23(1):63-7
12061719
J Biomol NMR. 2001 Dec;21(4):361-6
11824755
Science. 2003 Feb 28;299(5611):1362-7
12610298
Proc Natl Acad Sci U S A. 1996 Jan 23;93(2):861-6
8570649
J Am Chem Soc. 2001 Feb 21;123(7):1416-24
11456715
J Am Chem Soc. 2004 Feb 11;126(5):1314-5
14759169
J Biomol NMR. 2001 Dec;21(4):321-33
11824752
J Mol Biol. 1999 Jul 2;290(1):213-28
10388568
Annu Rev Genet. 1996;30:405-39
8982460
Science. 1993 Jun 4;260(5113):1491-6
8502992
J Biomol NMR. 2002 Nov;24(3):263-8
12522313
Chem Rev. 2004 Aug;104(8):3623-40
15303831
J Am Chem Soc. 2001 Jun 27;123(25):6098-107
11414844
Methods Enzymol. 2001;339:204-38
11462813
Biochemistry. 1992 Apr 14;31(14):3645-52
1314645
Structure. 2001 Oct;9(10):897-904
11591345
Proc Natl Acad Sci U S A. 2001 Jul 3;98(14):7684-9
11438724
J Mol Biol. 1999 Sep 3;291(5):1067-77
10518943
J Biomol NMR. 2000 Sep;18(1):33-42
11061226
FASEB J. 1997 Dec;11(14):1257-68
9409544
Methods Enzymol. 2001;338:3-34
11460554
J Magn Reson. 1998 Apr;131(2):351-7
9571112
J Am Chem Soc. 2004 Feb 25;126(7):2247-56
14971961
J Biomol NMR. 2004 Mar;28(3):263-72
14752259
Biopolymers. 2002 Dec 15;65(6):408-23
12434429
J Magn Reson. 1999 Jan;136(1):118-26
9887297