Probing structural transitions in the intrinsically disordered C-terminal domain of the measles virus nucleoprotein by vibrational spectroscopy of cyanylated cysteines.
Four single-cysteine variants of the intrinsically disordered C-terminal domain of the measles virus nucleoprotein (N(TAIL)) were cyanylated at cysteine and their infrared spectra in the C triple bond N stretching region were recorded both in the absence and in the presence of one of the physiological partners of N(TAIL), namely the C-terminal X domain (XD) of the viral phosphoprotein. Consistent with previous studies showing that XD triggers a disorder-to-order transition within N(TAIL), the C triple bond N stretching bands of the infrared probe were found to be significantly affected by XD, with this effect being position-dependent. When the cyanylated cysteine side chain is solvent-exposed throughout the structural transition, its changing linewidth reflects a local gain of structure. When the probe becomes partially buried due to binding, its frequency reports on the mean hydrophobicity of the microenvironment surrounding the labeled side chain of the bound form. The probe moiety is small compared to other common covalently attached spectroscopic probes, thereby minimizing possible steric hindrance/perturbation at the binding interface. These results show for the first time to our knowledge the suitability of site-specific cysteine mutagenesis followed by cyanylation and infrared spectroscopy to document structural transitions occurring within intrinsically disordered regions, with regions involved in binding and folding being identifiable at the residue level.
Bischak
Connor G
CG
Department of Chemistry, Haverford College, Haverford, Pennsylvania, USA.
Longhi
Sonia
S
Snead
David M
DM
Costanzo
Stéphanie
S
Terrer
Elodie
E
Londergan
Casey H
CH
D001665:Binding Sites; D003545:Cysteine / Q000737:chemistry*; D008459:Measles virus; D008958:Models, Molecular; D009570:Nitriles / Q000737:chemistry*; D009698:Nucleoproteins / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D013055:Spectrophotometry, Infrared / Q000379:methods*; D013379:Substrate Specificity; D014732:Vibration; D014764:Viral Proteins / Q000737:chemistry*
D009570:Nitriles; D009698:Nucleoproteins; D014764:Viral Proteins; D003545:Cysteine
Curr Opin Struct Biol. 2005 Feb;15(1):35-41
15718131
J Am Chem Soc. 2006 Oct 18;128(41):13356-7
17031938
J Phys Chem B. 2008 Aug 21;112(33):10352-7
18671422
J Am Chem Soc. 2003 Jan 15;125(2):405-11
12517152
Virus Res. 2004 Feb;99(2):157-67
14749181
J Am Chem Soc. 1970 Nov 18;92(23):6969-71
5483072
J Virol. 2004 Aug;78(16):8630-40
15280472
J Phys Chem B. 2010 Apr 15;114(14):4931-6
20297787
FEBS Lett. 2009 Apr 2;583(7):1084-9
19275899
Biophys J. 2010 Mar 17;98(6):1055-64
20303863
Nat Chem Biol. 2009 Jun;5(6):397-9
19396177
Nat Rev Mol Cell Biol. 2005 Mar;6(3):197-208
15738986
J Biol Chem. 2003 Nov 7;278(45):44567-73
12944395
Protein Pept Lett. 2010 Aug;17(8):961-78
20450481
J Mol Biol. 1999 Oct 22;293(2):321-31
10550212
Trends Biochem Sci. 2002 Jun;27(6):288-95
12069788
Curr Opin Struct Biol. 2009 Feb;19(1):23-30
19162471
Chem Phys Lett. 2009 Sep 1;478(4):249-253
20161057
J Chem Phys. 2008 Apr 21;128(15):154504
18433232
J Chem Phys. 2008 Apr 7;128(13):134506
18397076
Curr Opin Struct Biol. 2002 Feb;12(1):54-60
11839490
Nat Biotechnol. 2001 Sep;19(9):805-6
11533628
J Phys Chem A. 2007 Oct 11;111(40):10020-5
17867661
J Phys Chem Lett. 2010 Mar 4;1(5):850-855
20228945
J Biol Chem. 2002 Apr 5;277(14):11970-8
11812782
Curr Opin Struct Biol. 2009 Feb;19(1):31-8
19157855
Proteins. 2006 Jan 1;62(1):24-45
16287116
Pac Symp Biocomput. 1998;:473-84
9697205
J Phys Chem B. 2006 Oct 19;110(41):20596-608
17034249
Adv Protein Chem. 2002;62:25-49
12418100
J Mol Recognit. 2010 Sep-Oct;23(5):435-47
20058326
Protein Sci. 2005 Aug;14(8):1975-92
16046624
J Mol Recognit. 2005 Sep-Oct;18(5):343-84
16094605
Biochemistry. 1992 Sep 1;31(34):7922-30
1510978
Biochemistry. 2000 Jul 25;39(29):8396-405
10913245
Q Rev Biophys. 2007 Feb;40(1):1-53
17565764
Biochemistry. 1974 Jan 1;13(1):1-11
4808702
J Biol Chem. 2003 May 16;278(20):18638-48
12621042
Trends Biochem Sci. 2003 Feb;28(2):81-5
12575995
J Bioinform Comput Biol. 2006 Apr;4(2):597-608
16819805
Proteins. 2008 Dec;73(4):973-88
18536007
J Mol Graph Model. 2001;19(1):26-59
11381529
Virology. 2006 Jan 5;344(1):94-110
16364741