Cysteine misincorporation in bacterially expressed human alpha-synuclein - PubMed (original) (raw)
. 2006 Mar 20;580(7):1775-9.
doi: 10.1016/j.febslet.2006.02.032. Epub 2006 Feb 24.
Affiliations
- PMID: 16513114
- DOI: 10.1016/j.febslet.2006.02.032
Free article
Cysteine misincorporation in bacterially expressed human alpha-synuclein
Masami Masuda et al. FEBS Lett. 2006.
Free article
Abstract
Bacterially expressed human alpha-synuclein (alpha-syn) has been widely used in structural and functional studies. Here we show that approximately 20% of human alpha-syn expressed in Escherichia coli is mistranslated and that a Cys residue is incorporated at position 136 instead of a Tyr. Site-directed mutagenesis of codon 136 (TAC to TAT) resulted in the expression of alpha-syn lacking Cys. Although wild-type (Y136-TAC and Y136-TAT) and mutant (C136-TGC) alpha-syn had similar propensities to assemble into filaments, the levels of dimeric alpha-syn were increased by misincorporation. To avoid potential artefacts, we recommend use of the Y136-TAT construct for the expression of human alpha-syn.
Similar articles
- Dimerization of Tyr136Cys alpha-synuclein prevents amyloid transformation of wild type alpha-synuclein.
Barinova KV, Kuravsky ML, Arutyunyan AM, Serebryakova MV, Schmalhausen EV, Muronetz VI. Barinova KV, et al. Int J Biol Macromol. 2017 Mar;96:35-43. doi: 10.1016/j.ijbiomac.2016.12.011. Epub 2016 Dec 6. Int J Biol Macromol. 2017. PMID: 27939273 - Sequence- and seed-structure-dependent polymorphic fibrils of alpha-synuclein.
Tanaka G, Yamanaka T, Furukawa Y, Kajimura N, Mitsuoka K, Nukina N. Tanaka G, et al. Biochim Biophys Acta Mol Basis Dis. 2019 Jun 1;1865(6):1410-1420. doi: 10.1016/j.bbadis.2019.02.013. Epub 2019 Feb 18. Biochim Biophys Acta Mol Basis Dis. 2019. PMID: 30790619 - α-Synuclein in central nervous system and from erythrocytes, mammalian cells, and Escherichia coli exists predominantly as disordered monomer.
Fauvet B, Mbefo MK, Fares MB, Desobry C, Michael S, Ardah MT, Tsika E, Coune P, Prudent M, Lion N, Eliezer D, Moore DJ, Schneider B, Aebischer P, El-Agnaf OM, Masliah E, Lashuel HA. Fauvet B, et al. J Biol Chem. 2012 May 4;287(19):15345-64. doi: 10.1074/jbc.M111.318949. Epub 2012 Feb 7. J Biol Chem. 2012. PMID: 22315227 Free PMC article. - Tyrosine-to-cysteine modification of human alpha-synuclein enhances protein aggregation and cellular toxicity.
Zhou W, Freed CR. Zhou W, et al. J Biol Chem. 2004 Mar 12;279(11):10128-35. doi: 10.1074/jbc.M307563200. Epub 2003 Dec 29. J Biol Chem. 2004. PMID: 14699135 - Potential Effect of Post-Transcriptional Substitutions of Tyrosine for Cysteine Residues on Transformation of Amyloidogenic Proteins.
Muronetz VI, Pozdyshev DV, Medvedeva MV, Sevostyanova IA. Muronetz VI, et al. Biochemistry (Mosc). 2022 Feb;87(2):170-178. doi: 10.1134/S0006297922020080. Biochemistry (Mosc). 2022. PMID: 35508908 Review.
Cited by
- Lysosomal stress drives the release of pathogenic α-synuclein from macrophage lineage cells via the LRRK2-Rab10 pathway.
Abe T, Kuwahara T, Suenaga S, Sakurai M, Takatori S, Iwatsubo T. Abe T, et al. iScience. 2024 Jan 12;27(2):108893. doi: 10.1016/j.isci.2024.108893. eCollection 2024 Feb 16. iScience. 2024. PMID: 38313055 Free PMC article. - Molecular basis for preventing α-synuclein aggregation by a molecular tweezer.
Acharya S, Safaie BM, Wongkongkathep P, Ivanova MI, Attar A, Klärner FG, Schrader T, Loo JA, Bitan G, Lapidus LJ. Acharya S, et al. J Biol Chem. 2014 Apr 11;289(15):10727-10737. doi: 10.1074/jbc.M113.524520. Epub 2014 Feb 24. J Biol Chem. 2014. PMID: 24567327 Free PMC article. - The Wnt/Ca2+ pathway is involved in interneuronal communication mediated by tunneling nanotubes.
Vargas JY, Loria F, Wu YJ, Córdova G, Nonaka T, Bellow S, Syan S, Hasegawa M, van Woerden GM, Trollet C, Zurzolo C. Vargas JY, et al. EMBO J. 2019 Dec 2;38(23):e101230. doi: 10.15252/embj.2018101230. Epub 2019 Oct 18. EMBO J. 2019. PMID: 31625188 Free PMC article. - Single molecule characterization of α-synuclein in aggregation-prone states.
Trexler AJ, Rhoades E. Trexler AJ, et al. Biophys J. 2010 Nov 3;99(9):3048-55. doi: 10.1016/j.bpj.2010.08.056. Biophys J. 2010. PMID: 21044603 Free PMC article. - Allostery in a disordered protein: oxidative modifications to α-synuclein act distally to regulate membrane binding.
Sevcsik E, Trexler AJ, Dunn JM, Rhoades E. Sevcsik E, et al. J Am Chem Soc. 2011 May 11;133(18):7152-8. doi: 10.1021/ja2009554. Epub 2011 Apr 14. J Am Chem Soc. 2011. PMID: 21491910 Free PMC article.
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Miscellaneous