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Assay of Arginyltransferase Activity by a Fluorescent HPLC Method

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Protein Arginylation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1337))

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Abstract

Syntheses of fluorescent substrate and product for arginyltransferase, N-aspartyl-N′-dansylamido-1,4-butanediamine (Asp-4DNS), and N-arginylaspartyl-N′-dansylamido-1,4-butanediamine (ArgAsp-4DNS), respectively, including their precursor 4-dansylamidobutylamine (4DNS), are described. Then, HPLC conditions are summarized for a baseline separation of the three compounds in 10 min. The present method, which permits the simultaneous determination of Asp-4DNS, 4DNS, and ArgAsp-4DNS (in eluting order), is advantageous in measuring arginyltransferase activity and detecting the unfavorable enzyme(s) in 105,000 × g supernatant of tissues to ensure accurate determination.

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Acknowledgments

The author would like to thank Dr. Keijiro Samejima for his help with this manuscript.

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Correspondence to Koichi Takao .

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Takao, K. (2015). Assay of Arginyltransferase Activity by a Fluorescent HPLC Method. In: Kashina, A. (eds) Protein Arginylation. Methods in Molecular Biology, vol 1337. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2935-1_12

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  • DOI: https://doi.org/10.1007/978-1-4939-2935-1_12

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2934-4

  • Online ISBN: 978-1-4939-2935-1

  • eBook Packages: Springer Protocols

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