Role of Tyr306 in the C-terminal fragment of Clostridium perfringens enterotoxin for modulation of tight junction |
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Authors: | Ebihara Chiaki Kondoh Masuo Harada Motoki Fujii Makiko Mizuguchi Hiroyuki Tsunoda Shin-Ichi Horiguchi Yasuhiko Yagi Kiyohito Watanabe Yoshiteru |
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Affiliation: | Department of Pharmaceutics and Biopharmaceutics, Showa Pharmaceutical University, Machida, Tokyo 194-8543, Japan. |
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Abstract: | We previously reported that the C-terminal fragment of Clostridium perfringens enterotoxin (C-CPE) is a novel type of absorption enhancer that interacts with claudin-4 and that Tyr306 of C-CPE plays a role in ability of C-CPE to modulate barrier of tight junctions. In the current study, to investigate effects of Tyr306 on the C-CPE activity, we prepared some C-CPE mutants substituted Tyr306 with Trp (Y306W), Phe (Y306F) and Lys (Y306K). We found that Y306W and Y306F mutants of C-CPE had claudin-4 binding affinities and effects on the barrier function of tight junctions, whereas both of these properties were greatly reduced with the Y306K mutant. Finally, the Y306K but not the Y306F and Y306W mutants had reduced abilities to enhance absorption in rat jejunum. These results indicate that aromatic and hydrophobic properties, not hydrogen bonding potential, of Tyr306 are involved in the interaction of C-CPE with claudin-4 and in the modulation of the tight junction barrier function by C-CPE. |
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Keywords: | C-CPE, the C-terminal fragment of Clostridium perfringens enterotoxin PSIF, protein synthesis inhibitory factor TJ, tight junction CPE, Clostridium perfringens enterotoxin TER, transepithelial electric resistance C-CPE-PSIF, C-CPE fused to PSIF PCR, polymerase chain reaction LDH, lactate dehydrogenase FD-4, fluorescein-isothiocyanate-dextran with a molecular weight of 4000 |
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