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Development of New Tools for the Studies of Protein Arginylation

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Protein Arginylation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2620))

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Abstract

Studies of posttranslational modifications present many unique challenges, stemming from their role as the major drivers of biological complexity. Perhaps the most immediate challenge to researchers working on virtually any posttranslational modification is the shortage of reliable easy-to-use tools that can enable massive identification and characterization of posttranslationally modified proteins, as well as their functional modulation in vitro and in vivo. In the case of protein arginylation, which utilizes charged Arg-tRNA that is also used by the ribosomes, detection and labeling of arginylated proteins is especially difficult, because of the necessity of distinguishing these proteins from the products of conventional translation. As of now, this difficulty remains the major obstacle to new researchers entering the field. This chapter discusses some of the strategies for developing antibodies for arginylation detection, as well as some general considerations for development of other tools for studies of arginylation.

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References

  1. Wong CCL, Xu T, Rai R, Bailey AO, Yates JR, Wolf YI, Zebroski H, Kashina A (2007) Global analysis of posttranslational protein Arginylation. PLoS Biol 5(10):e258

    Article  PubMed  PubMed Central  Google Scholar 

  2. Wang J, Han X, Saha S, Xu T, Rai R, Zhang F, Wolf YI, Wolfson A, Yates JR 3rd, Kashina A (2011) Arginyltransferase is an ATP-independent self-regulating enzyme that forms distinct functional complexes in vivo. Chem Biol 18(1):121–130. https://doi.org/10.1016/j.chembiol.2010.10.016

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Wang J, Han X, Wong CC, Cheng H, Aslanian A, Xu T, Leavis P, Roder H, Hedstrom L, Yates JR 3rd, Kashina A (2014) Arginyltransferase ATE1 catalyzes midchain Arginylation of proteins at side chain carboxylates in vivo. Chem Biol 21(3):331–337. https://doi.org/10.1016/j.chembiol.2013.12.017

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Leibowitz MJ, Soffer RL (1969) A soluble enzyme from Escherichia coli which catalyzes the transfer of leucine and phenylalanine from tRNA to acceptor proteins. Biochem Biophys Res Commun 36(1):47–53

    Article  CAS  PubMed  Google Scholar 

  5. Wagner AM, Fegley MW, Warner JB, Grindley CLJ, Marotta NP, Petersson EJ (2011) N-terminal protein modification using simple aminoacyl transferase substrates. J Am Chem Soc 133(38):15139–15147. https://doi.org/10.1021/ja2055098

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Tanaka T, Wagner AM, Warner JB, Wang YJ, Petersson EJ (2013) Expressed protein ligation at methionine: N-terminal attachment of homocysteine, ligation, and masking. Angew Chem Int Ed Engl 52(24):6210–6213. https://doi.org/10.1002/anie.201302065

    Article  CAS  PubMed  Google Scholar 

  7. Avcilar-Kucukgoze I, Gamper H, Polte C, Ignatova Z, Kraetzner R, Shtutman M, Hou Y-M, Dong DW, Kashina A (2020) tRNAArg-derived fragments can serve as Arginine donors for protein arginylation. Cell Chem Biol 27(7):839–849.e4 S2451945620301902. https://doi.org/10.1016/j.chembiol.2020.05.013

  8. Van V, Ejimogu N-E, Bui TS, Smith AT (2022) The Structure of Saccharomyces cerevisiae Arginyltransferase 1 (ATE1). J Mol Biol 434(21):167816-S0022283622004351. https://doi.org/10.1016/j.jmb.2022.167816

  9. Kim BH, Kim MK, Oh SJ, Nguyen KT, Kim JH, Varshavsky A, Hwang C-S, Song HK (2022) Crystal structure of the Ate1 arginyl-tRNA-protein transferase and arginylation of N-degron substrates. Proc Natl Acad Sci 119(31):e2209597119. https://doi.org/10.1073/pnas.2209597119

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Correspondence to Anna S. Kashina .

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Kashina, A.S. (2023). Development of New Tools for the Studies of Protein Arginylation. In: Kashina, A.S. (eds) Protein Arginylation. Methods in Molecular Biology, vol 2620. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2942-0_30

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  • DOI: https://doi.org/10.1007/978-1-0716-2942-0_30

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2941-3

  • Online ISBN: 978-1-0716-2942-0

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