Abstract
The location of disulfide linkage(s) or status of unpaired cysteines is a critical structural feature required for the characterization of three-dimensional structure of a protein and for the correlation of protein structure–function relationships. Cysteine, with its reactive thiol group, can undergo enzymatic or oxidative posttranslational modification in response to changing redox conditions to signal a cascade of downstream reactions. In such a situation, it becomes even more critical to obtain the information on the pair of cysteines involved in such a redox switch operation. Here, a method involving chemical derivatization and liquid chromatography–mass spectrometry (LC-MS) is described to determine the cysteine residues involved in disulfide bond formation for a protein containing multiple cysteines in its sequence.
Key words
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Gilmartin GM (2005) Eukaryotic mRNA 3′ processing: a common means to different ends. Genes Dev 19(21):2517–2521. doi:10.1101/gad.1378105
Hunt AG, Xing D, Li QQ (2012) Plant polyadenylation factors: conservation and variety in the polyadenylation complex in plants. BMC Genomics 13:641. doi:10.1186/1471-2164-13-641
Hunt AG, Xu R, Addepalli B, Rao S, Forbes KP, Meeks LR, Xing D, Mo M, Zhao H, Bandyopadhyay A, Dampanaboina L, Marion A, Von Lanken C, Li QQ (2008) Arabidopsis mRNA polyadenylation machinery: comprehensive analysis of protein-protein interactions and gene expression profiling. BMC Genomics 9:220. doi:10.1186/1471-2164-9-220
Mandel CR, Bai Y, Tong L (2008) Protein factors in pre-mRNA 3′-end processing. Cell Mol Life Sci 65(7–8):1099–1122. doi:10.1007/s00018-007-7474-3
Zhang J, Addepalli B, Yun KY, Hunt AG, Xu R, Rao S, Li QQ, Falcone DL (2008) A polyadenylation factor subunit implicated in regulating oxidative signaling in Arabidopsis thaliana. PLoS One 3(6):e2410. doi:10.1371/journal.pone.0002410
Buchanan BB, Balmer Y (2005) Redox regulation: a broadening horizon. Annu Rev Plant Biol 56:187–220. doi:10.1146/annurev.arplant.56.032604.144246
Bykova NV, Rampitsch C (2013) Modulating protein function through reversible oxidation: redox-mediated processes in plants revealed through proteomics. Proteomics 13(3–4):579–596. doi:10.1002/pmic.201200270
Paulsen CE, Carroll KS (2010) Orchestrating redox signaling networks through regulatory cysteine switches. ACS Chem Biol 5(1):47–62. doi:10.1021/cb900258z
Winterbourn CC (2008) Reconciling the chemistry and biology of reactive oxygen species. Nat Chem Biol 4(5):278–286. doi:10.1038/nchembio.85
Murray CI, Van Eyk JE (2012) Chasing cysteine oxidative modifications: proteomic tools for characterizing cysteine redox status. Circ Cardiovasc Genet 5(5):591. doi:10.1161/CIRCGENETICS.111.961425
Bachi A, Dalle-Donne I, Scaloni A (2013) Redox proteomics: chemical principles, methodological approaches and biological/biomedical promises. Chem Rev 113(1):596–698. doi:10.1021/cr300073p
Chouchani ET, James AM, Fearnley IM, Lilley KS, Murphy MP (2011) Proteomic approaches to the characterization of protein thiol modification. Curr Opin Chem Biol 15(1):120–128. doi:10.1016/j.cbpa.2010.11.003
Han B, Hare M, Wickramasekara S, Fang Y, Maier CS (2012) A comparative ‘bottom up’ proteomics strategy for the site-specific identification and quantification of protein modifications by electrophilic lipids. J Proteomics 75(18):5724–5733. doi:10.1016/j.jprot.2012.07.029
Izquierdo-Alvarez A, Martinez-Ruiz A (2011) Thiol redox proteomics seen with fluorescent eyes: the detection of cysteine oxidative modifications by fluorescence derivatization and 2-DE. J Proteomics 75(2):329–338. doi:10.1016/j.jprot.2011.09.013
Gallegos-Perez JL, Rangel-Ordonez L, Bowman SR, Ngowe CO, Watson JT (2005) Study of primary amines for nucleophilic cleavage of cyanylated cystinyl proteins in disulfide mass mapping methodology. Anal Biochem 346(2):311–319. doi:10.1016/j.ab.2005.08.003
Wefing S, Schnaible V, Hoffmann D (2006) SearchXLinks. A program for the identification of disulfide bonds in proteins from mass spectra. Anal Chem 78(4):1235–1241. doi:10.1021/ac051634x
Bach RD, Dmitrenko O, Thorpe C (2008) Mechanism of thiolate-disulfide interchange reactions in biochemistry. J Org Chem 73(1):12–21. doi:10.1021/jo702051f
Addepalli B, Hunt AG (2008) Ribonuclease activity is a common property of Arabidopsis CCCH-containing zinc-finger proteins. FEBS Lett 582(17):2577–2582. doi:10.1016/j.febslet.2008.06.029
Addepalli B, Limbach PA, Hunt AG (2010) A disulfide linkage in a CCCH zinc finger motif of an Arabidopsis CPSF30 ortholog. FEBS Lett 584(21):4408–4412. doi:10.1016/j.febslet.2010.09.043
Eng JK, McCormack AL, Yates Iii JR (1994) An approach to correlate tandem mass spectral data of peptides with amino acid sequences in a protein database. J Am Soc Mass Spectrom 5(11):976–989. doi:10.1016/1044-0305(94)80016-2
Cottrell JS (2011) Protein identification using MS/MS data. J Proteomics 74(10):1842–1851. doi:10.1016/j.jprot.2011.05.014
Mo J, Tymiak AA, Chen G (2013) Characterization of disulfide linkages in recombinant human granulocyte-colony stimulating factor. Rapid Commun Mass Spectrom 27(9):940–946. doi:10.1002/rcm.6530
Wu SL, Jiang H, Lu Q, Dai S, Hancock WS, Karger BL (2009) Mass spectrometric determination of disulfide linkages in recombinant therapeutic proteins using online LC-MS with electron-transfer dissociation. Anal Chem 81(1):112–122. doi:10.1021/ac801560k
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Addepalli, B. (2015). Detection of Disulfide Linkage by Chemical Derivatization and Mass Spectrometry. In: Hunt, A., Li, Q. (eds) Polyadenylation in Plants. Methods in Molecular Biology, vol 1255. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2175-1_10
Download citation
DOI: https://doi.org/10.1007/978-1-4939-2175-1_10
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2174-4
Online ISBN: 978-1-4939-2175-1
eBook Packages: Springer Protocols