Pillar[6]arene acts as a biosensor for quantitative detection of a vitamin metabolite in crude biological samples

Masaya Ueno; Takuya Tomita; Hiroshi Arakawa; Takahiro Kakuta; Tada-Aki Yamagishi; Jumpei Terakawa; Takiko Daikoku; Shin-Ichi Horike; Sha Si; Kenta Kurayoshi; Chiaki Ito; Atsuko Kasahara; Yuko Tadokoro; Masahiko Kobayashi; Tsutomu Fukuwatari; Ikumi Tamai; Atsushi Hirao; Tomoki Ogoshi

doi:10.1038/s42004-020-00430-w

Pillar[6]arene acts as a biosensor for quantitative detection of a vitamin metabolite in crude biological samples

Commun Chem. 2020 Dec 7;3(1):183. doi: 10.1038/s42004-020-00430-w.

Authors

Masaya Ueno^#^{1

2}, Takuya Tomita^#³, Hiroshi Arakawa⁴, Takahiro Kakuta^{2

3}, Tada-Aki Yamagishi³, Jumpei Terakawa⁵, Takiko Daikoku⁵, Shin-Ichi Horike⁶, Sha Si^{1

2}, Kenta Kurayoshi¹, Chiaki Ito¹, Atsuko Kasahara⁷, Yuko Tadokoro^{1

2}, Masahiko Kobayashi^{1

2}, Tsutomu Fukuwatari⁸, Ikumi Tamai⁴, Atsushi Hirao^{9

10}, Tomoki Ogoshi^{11

12}

Affiliations

¹ Division of Molecular Genetics, Cancer and Stem Cell Research Program, Cancer Research Institute, Kanazawa University, Kakuma-machi, Kanazawa, Ishikawa, 920-1192, Japan.
² WPI Nano Life Science Institute (WPI-Nano LSI), Kanazawa University, Kakuma-machi, Kanazawa, Ishikawa, 920-1192, Japan.
³ Graduate School of Natural Science and Technology, Kanazawa University, Kakuma-machi, Kanazawa, Ishikawa, 920-1192, Japan.
⁴ Faculty of Pharmaceutical Sciences, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Kakuma-machi, Kanazawa, Ishikawa, 920-1192, Japan.
⁵ Institute for Experimental Animals, Advanced Science Research Center, Kanazawa University, Takara-machi, Kanazawa, 920-8641, Japan.
⁶ Division of Functional Genomics, Advanced Science Research Center, Kanazawa University, Takara-machi, Kanazawa, 920-8641, Japan.
⁷ Institute for Frontier Science Initiative, Kanazawa University, Kakuma-machi, Kanazawa, Ishikawa, 920-1192, Japan.
⁸ Department of Nutrition, School of Human Cultures, The University of Shiga Prefecture, 2500 Hassaka, Hikone, Shiga, 522-8533, Japan.
⁹ Division of Molecular Genetics, Cancer and Stem Cell Research Program, Cancer Research Institute, Kanazawa University, Kakuma-machi, Kanazawa, Ishikawa, 920-1192, Japan. ahirao@staff.kanazawa-u.ac.jp.
¹⁰ WPI Nano Life Science Institute (WPI-Nano LSI), Kanazawa University, Kakuma-machi, Kanazawa, Ishikawa, 920-1192, Japan. ahirao@staff.kanazawa-u.ac.jp.
¹¹ WPI Nano Life Science Institute (WPI-Nano LSI), Kanazawa University, Kakuma-machi, Kanazawa, Ishikawa, 920-1192, Japan. ogoshi@sbchem.kyoto-u.ac.jp.
¹² Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering Kyoto University, Kyoto, 615-8510, Japan. ogoshi@sbchem.kyoto-u.ac.jp.

^# Contributed equally.

Abstract

Metabolic syndrome is associated with obesity, hypertension, and dyslipidemia, and increased cardiovascular risk. Therefore, quick and accurate measurements of specific metabolites are critical for diagnosis; however, detection methods are limited. Here we describe the synthesis of pillar[n]arenes to target 1-methylnicotinamide (1-MNA), which is one metabolite of vitamin B3 (nicotinamide) produced by the cancer-associated nicotinamide N-methyltransferase (NNMT). We found that water-soluble pillar[5]arene (P5A) forms host-guest complexes with both 1-MNA and nicotinamide, and water-soluble pillar[6]arene (P6A) selectively binds to 1-MNA at the micromolar level. P6A can be used as a "turn-off sensor" by photoinduced electron transfer (detection limit is 4.38 × 10^-6 M). In our cell-free reaction, P6A is used to quantitatively monitor the activity of NNMT. Moreover, studies using NNMT-deficient mice reveal that P6A exclusively binds to 1-MNA in crude urinary samples. Our findings demonstrate that P6A can be used as a biosensor to quantify 1-MNA in crude biological samples.

Abstract

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