Production and characterization of recombinant tachycitin, the Cys-rich chitin-binding protein
| Autor: | Tsukasa Osaki, Yoshihiro Kobashigawa, Nozomi Koganesawa, Katsutoshi Nitta, Keiichi Kawano, Sakae Tsuda, Tetsuya Suetake, Shun Ichiro Kawabata, Tomoyasu Aizawa, Makoto Demura |
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| Rok vydání: | 2002 |
| Předmět: |
Models
Molecular Protein Conformation Bioengineering Chitin Biology In Vitro Techniques Protein Engineering Biochemistry law.invention chemistry.chemical_compound law Chitin binding Amide Gene expression Animals Amino Acid Sequence Cysteine Molecular Biology Nuclear Magnetic Resonance Biomolecular Binding Sites Base Sequence Blood Proteins DNA biology.organism_classification Antimicrobial Protein tertiary structure Recombinant Proteins Anti-Bacterial Agents chemistry Recombinant DNA Carrier Proteins Bacteria Biotechnology |
| Zdroj: | Protein engineering. 15(9) |
| ISSN: | 0269-2139 |
| Popis: | Tachycitin is an invertebrate chitin-binding protein with an amidated C-terminus, and possesses antimicrobial activity against both fungi and bacteria. The 1 H-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, 1 5 N-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 1 5 N 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. |
| Databáze: | OpenAIRE |
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