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Inhibitory Effect on RT-PCR and Restriction Enzyme Activity by Ommochrome and Its Mechanism
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Abstract
To explore the physiological role and/or pharmacological effects of ommochrome, which is a natural organic pigment widely distributed in Protostomia, we attempted to investigate the influence of ommochrome on RT-PCR and activities of restriction enzymes. It was found that ommin, an ommochrome purified from the diapause eggs of Bombyx mori, inhibited the RT-PCR and restriction enzyme activities. The mechanism of these inhibitory reactions is assumed to be the direct binding of ommochrome to DNA rather than acting against the enzymes because, similarly to actinomycin D, there is a phenoxazine ring in the structure of ommin that is known to be intercalated to DNA. To reveal the ommin/DNA interaction, it was investigated by computational approaches such as molecular docking, molecular dynamics simulation, and free energy calculation. From the computational analyses, it was expected that ommin would bind to DNA with almost the same strength as actinomycin D and intercalate into DNA. This is the first report on the pharmacological effect of ommochrome and its inhibitory mechanism obtained from biochemical and computational analyses.
Received 07 7月 2023
Accepted 20 9月 2023
Acknowledgments:
We would like to thank Rina Chigasaki, Saya Abe, Misaki Okita, Saki Uematsu, Yoshio Shimamura, Kanako Fujikawa, Chisaki Ogawa, and Ayame Otsuki, students of Nihon University, for their help in some experiments. This work was supported by a Nihon University Research Grant from 2017 to 2023.
This paper is dedicated to the late Professor Motoo Tsusué and the late Dr. Toshio Yamamoto, the dear mentors of the corresponding author.
© 2023 Zoological Society of Japan
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