Production and characterization of recombinant tachycitin, the Cys-rich chitin-binding protein

Tetsuya Suetake, Tomoyasu Aizawa, Nozomi Koganesawa, Tsukasa Osaki, Yoshihiro Kobashigawa, Makoto Demura, Shun-Ichiro Kawabata, Keiichi Kawano, Sakae Tsuda, Katsutoshi Nitta

Research output: Contribution to journalArticle

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Abstract

Tachycitin is an invertebrate chitin-binding protein with an amidated C-terminus, and possesses antimicrobial activity against both fungi and bacteria. The 1H-NMR-based tertiary structure of tachycitin was recently determined [Suetake et al. (2000) J. Biol. Chem., 275, 17929-17932]. In order to examine the structural and functional features of tachycitin more closely, we performed for the first time, gene expression, refolding, 15N-NMR-based characterizations, and antimicrobial activity measurements of a recombinant tachycitin (rTcn) that does not have the amide group at the C-terminus. The NMR analysis indicated that rTcn possesses the same structural construction as the native tachycitin. The backbone 15N relaxation measurements showed that the molecular motional correlation time of rTcn increases as its concentration increases, indicating that tachycitins have a tendency to aggregate with each other. rTcn exhibits antimicrobial activity against fungi but not against bacteria. The cell surface of fungi contains chitin as an essential constituent, but that of bacteria does not. These results suggest that not only the chitin-binding region but also the C-terminal amide group of tachycitin plays a significant role in its antimicrobial properties.

Original languageEnglish
Pages (from-to)763-769
Number of pages7
JournalProtein Engineering
Volume15
Issue number9
Publication statusPublished - Sep 1 2002

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Chitin
Fungi
Bacteria
Carrier Proteins
Nuclear magnetic resonance
Amides
Invertebrates
Gene expression
Gene Expression

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Suetake, T., Aizawa, T., Koganesawa, N., Osaki, T., Kobashigawa, Y., Demura, M., ... Nitta, K. (2002). Production and characterization of recombinant tachycitin, the Cys-rich chitin-binding protein. Protein Engineering, 15(9), 763-769.

Production and characterization of recombinant tachycitin, the Cys-rich chitin-binding protein. / Suetake, Tetsuya; Aizawa, Tomoyasu; Koganesawa, Nozomi; Osaki, Tsukasa; Kobashigawa, Yoshihiro; Demura, Makoto; Kawabata, Shun-Ichiro; Kawano, Keiichi; Tsuda, Sakae; Nitta, Katsutoshi.

In: Protein Engineering, Vol. 15, No. 9, 01.09.2002, p. 763-769.

Research output: Contribution to journalArticle

Suetake, T, Aizawa, T, Koganesawa, N, Osaki, T, Kobashigawa, Y, Demura, M, Kawabata, S-I, Kawano, K, Tsuda, S & Nitta, K 2002, 'Production and characterization of recombinant tachycitin, the Cys-rich chitin-binding protein', Protein Engineering, vol. 15, no. 9, pp. 763-769.
Suetake T, Aizawa T, Koganesawa N, Osaki T, Kobashigawa Y, Demura M et al. Production and characterization of recombinant tachycitin, the Cys-rich chitin-binding protein. Protein Engineering. 2002 Sep 1;15(9):763-769.
Suetake, Tetsuya ; Aizawa, Tomoyasu ; Koganesawa, Nozomi ; Osaki, Tsukasa ; Kobashigawa, Yoshihiro ; Demura, Makoto ; Kawabata, Shun-Ichiro ; Kawano, Keiichi ; Tsuda, Sakae ; Nitta, Katsutoshi. / Production and characterization of recombinant tachycitin, the Cys-rich chitin-binding protein. In: Protein Engineering. 2002 ; Vol. 15, No. 9. pp. 763-769.
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