title: Highly sensitive detection of E2 activity in ubiquitination using an artificial RING finger.
abstract: The ubiquitin-conjugating (E2) enzymes of protein ubiquitination are associated with various diseases such as leukemia, lung cancer, and breast cancer. Rapid and accurate detection of E2 enzymatic activities remains poor. Here, we described the detection of E2 activity on a signal accumulation ISFET biosensor (AMIS sensor) using an artificial RING finger (ARF). The use of ARF enables the simplified detection of E2 activity without a substrate. The high-sensitivity quantitative detection of E2 activities was demonstrated via real-time monitoring over a response range of femtomolar to micromolar concentrations. Furthermore, the monitoring of E2 activities was successfully achieved using human acute promyelocytic leukemia cells following treatment with the anticancer drug bortezomib, which allowed the assessment of the pathological conditions. This strategy is extremely simple and convenient, and the present detection could be widely applied to specific E2s for various types of cancers. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.
authors.affiliation: Department of Pharmaceutical Health Care, Faculty of Pharmaceutical Sciences, Himeji Dokkyo University, Hyogo, Japan.
authors.forename: Kazuhide
authors.initials: K
authors.lastname: Miyamoto
authors.affiliation: Department of Pharmaceutical Health Care, Faculty of Pharmaceutical Sciences, Himeji Dokkyo University, Hyogo, Japan.
authors.forename: Miho
authors.initials: M
authors.lastname: Sumida
authors.affiliation: Department of Pharmaceutical Health Care, Faculty of Pharmaceutical Sciences, Himeji Dokkyo University, Hyogo, Japan.
authors.forename: Mayumi
authors.initials: M
authors.lastname: Yuasa-Sunagawa
authors.affiliation: Department of Pharmaceutical Health Care, Faculty of Pharmaceutical Sciences, Himeji Dokkyo University, Hyogo, Japan.
authors.forename: Kazuki
authors.initials: K
authors.lastname: Saito
mesh_terms: D000595:Amino Acid Sequence; D000970:Antineoplastic Agents / Q000494:pharmacology; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D000069286:Bortezomib / Q000494:pharmacology; D045744:Cell Line, Tumor; D055664:Electrochemical Techniques; D015973:Gene Expression Regulation, Leukemic; D042381:Granulocyte Precursor Cells / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D057786:Peptidomimetics / Q000138:chemical synthesis / Q000378:metabolism*; D011522:Protons; D054829:RING Finger Domains / Q000235:genetics; D015398:Signal Transduction; D060327:Solid-Phase Synthesis Techniques; D025801:Ubiquitin / Q000235:genetics / Q000378:metabolism; D044763:Ubiquitin-Conjugating Enzymes / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D054875:Ubiquitination / Q000187:drug effects
chemical_list: D000970:Antineoplastic Agents; D057786:Peptidomimetics; D011522:Protons; D025801:Ubiquitin; D000069286:Bortezomib; D044763:Ubiquitin-Conjugating Enzymes
keywords: E2 activity; artificial RING finger; biosensor; cancer; diagnosis; ubiquitination