title: Probing structural transitions in the intrinsically disordered C-terminal domain of the measles virus nucleoprotein by vibrational spectroscopy of cyanylated cysteines.
abstract: 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.
authors.affiliation: Department of Chemistry, Haverford College, Haverford, Pennsylvania, USA.
authors.forename: Connor G
authors.initials: CG
authors.lastname: Bischak
authors.forename: Sonia
authors.initials: S
authors.lastname: Longhi
authors.forename: David M
authors.initials: DM
authors.lastname: Snead
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Costanzo
authors.forename: Elodie
authors.initials: E
authors.lastname: Terrer
authors.forename: Casey H
authors.initials: CH
authors.lastname: Londergan
mesh_terms: 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*
chemical_list: D009570:Nitriles; D009698:Nucleoproteins; D014764:Viral Proteins; D003545:Cysteine
references.citation: Curr Opin Struct Biol. 2005 Feb;15(1):35-41
references.pmid: 15718131
references.citation: J Am Chem Soc. 2006 Oct 18;128(41):13356-7
references.pmid: 17031938
references.citation: J Phys Chem B. 2008 Aug 21;112(33):10352-7
references.pmid: 18671422
references.citation: J Am Chem Soc. 2003 Jan 15;125(2):405-11
references.pmid: 12517152
references.citation: Virus Res. 2004 Feb;99(2):157-67
references.pmid: 14749181
references.citation: J Am Chem Soc. 1970 Nov 18;92(23):6969-71
references.pmid: 5483072
references.citation: J Virol. 2004 Aug;78(16):8630-40
references.pmid: 15280472
references.citation: J Phys Chem B. 2010 Apr 15;114(14):4931-6
references.pmid: 20297787
references.citation: FEBS Lett. 2009 Apr 2;583(7):1084-9
references.pmid: 19275899
references.citation: Biophys J. 2010 Mar 17;98(6):1055-64
references.pmid: 20303863
references.citation: Nat Chem Biol. 2009 Jun;5(6):397-9
references.pmid: 19396177
references.citation: Nat Rev Mol Cell Biol. 2005 Mar;6(3):197-208
references.pmid: 15738986
references.citation: J Biol Chem. 2003 Nov 7;278(45):44567-73
references.pmid: 12944395
references.citation: Protein Pept Lett. 2010 Aug;17(8):961-78
references.pmid: 20450481
references.citation: J Mol Biol. 1999 Oct 22;293(2):321-31
references.pmid: 10550212
references.citation: Trends Biochem Sci. 2002 Jun;27(6):288-95
references.pmid: 12069788
references.citation: Curr Opin Struct Biol. 2009 Feb;19(1):23-30
references.pmid: 19162471
references.citation: Chem Phys Lett. 2009 Sep 1;478(4):249-253
references.pmid: 20161057
references.citation: J Chem Phys. 2008 Apr 21;128(15):154504
references.pmid: 18433232
references.citation: J Chem Phys. 2008 Apr 7;128(13):134506
references.pmid: 18397076
references.citation: Curr Opin Struct Biol. 2002 Feb;12(1):54-60
references.pmid: 11839490
references.citation: Nat Biotechnol. 2001 Sep;19(9):805-6
references.pmid: 11533628
references.citation: J Phys Chem A. 2007 Oct 11;111(40):10020-5
references.pmid: 17867661
references.citation: J Phys Chem Lett. 2010 Mar 4;1(5):850-855
references.pmid: 20228945
references.citation: J Biol Chem. 2002 Apr 5;277(14):11970-8
references.pmid: 11812782
references.citation: Curr Opin Struct Biol. 2009 Feb;19(1):31-8
references.pmid: 19157855
references.citation: Proteins. 2006 Jan 1;62(1):24-45
references.pmid: 16287116
references.citation: Pac Symp Biocomput. 1998;:473-84
references.pmid: 9697205
references.citation: J Phys Chem B. 2006 Oct 19;110(41):20596-608
references.pmid: 17034249
references.citation: Adv Protein Chem. 2002;62:25-49
references.pmid: 12418100
references.citation: J Mol Recognit. 2010 Sep-Oct;23(5):435-47
references.pmid: 20058326
references.citation: Protein Sci. 2005 Aug;14(8):1975-92
references.pmid: 16046624
references.citation: J Mol Recognit. 2005 Sep-Oct;18(5):343-84
references.pmid: 16094605
references.citation: Biochemistry. 1992 Sep 1;31(34):7922-30
references.pmid: 1510978
references.citation: Biochemistry. 2000 Jul 25;39(29):8396-405
references.pmid: 10913245
references.citation: Q Rev Biophys. 2007 Feb;40(1):1-53
references.pmid: 17565764
references.citation: Biochemistry. 1974 Jan 1;13(1):1-11
references.pmid: 4808702
references.citation: J Biol Chem. 2003 May 16;278(20):18638-48
references.pmid: 12621042
references.citation: Trends Biochem Sci. 2003 Feb;28(2):81-5
references.pmid: 12575995
references.citation: J Bioinform Comput Biol. 2006 Apr;4(2):597-608
references.pmid: 16819805
references.citation: Proteins. 2008 Dec;73(4):973-88
references.pmid: 18536007
references.citation: J Mol Graph Model. 2001;19(1):26-59
references.pmid: 11381529
references.citation: Virology. 2006 Jan 5;344(1):94-110
references.pmid: 16364741