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Design of a System for Monitoring Ubiquitination Activities of E2 Enzymes Using Engineered RING Finger Proteins

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Zinc Finger Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1867))

Abstract

Ubiquitination is a sequential cascade consisting of ubiquitin-activating (E1), ubiquitin-conjugating (E2), and ubiquitin-ligating (E3) enzymes. It controls numerous processes such as protein degradation, DNA repair, and signal transduction pathways. E2 enzymes are associated with a variety of diseases such as leukemia, breast cancer, lung cancer, and colorectal cancer. To date, the monitoring of E2 activity for cancer diagnosis is challenging due to its intricate cascade reaction. To surmount this hurdle, we have recently developed a novel strategy for monitoring E2 activities. Here, we describe the concise machinery of ubiquitination with artificial RING finger proteins (ARFs) functioning as E3 enzymes. This machinery enables the simplified monitoring of E2 activities. Furthermore, our system combines a signal accumulation ion-sensitive field-effect transistor biosensor with ARFs, allowing for real-time monitoring of the pathological conditions of cancer cells. The present methodology may lead to novel diagnostic techniques for cancers.

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Acknowledgments

This research was supported by A-STEP from Japan Science and Technology Agency (JST), a Grant-in-Aid for Scientific Research (KAKENHI 26430147), Takeda Science Foundation, Sanyo Chemical Industries Foundation, and Nakatani Foundation.

Author contributions

K.M. designed this study. K.M. and K.S. wrote the main manuscript text and prepared all the figures.

Competing interests

The authors declare no competing financial interests.

Author information

Authors and Affiliations

  1. Faculty of Pharmaceutical Sciences, Department of Pharmaceutical Health Care, Himeji Dokkyo University, Hyogo, Japan

    Kazuhide Miyamoto & Kazuki Saito

Authors
  1. Kazuhide Miyamoto
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  2. Kazuki Saito
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Corresponding author

Correspondence to Kazuhide Miyamoto .

Editor information

Editors and Affiliations

  1. Shanghai Institute for Advanced Immunochemical Studies, ShanghaiTech University, Shanghai, China

    Jia Liu

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Miyamoto, K., Saito, K. (2018). Design of a System for Monitoring Ubiquitination Activities of E2 Enzymes Using Engineered RING Finger Proteins. In: Liu, J. (eds) Zinc Finger Proteins. Methods in Molecular Biology, vol 1867. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8799-3_6

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  • DOI: https://doi.org/10.1007/978-1-4939-8799-3_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8798-6

  • Online ISBN: 978-1-4939-8799-3

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