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Conformational change in the active center region of GST P1-1, due to binding of a synthetic conjugate of DXR with GSH, enhanced JNK-mediated apoptosis

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Abstract

Treatment of cells with a synthetic conjugate of DXR with GSH via glutaraldehyde (GSH-DXR) caused cytochrome c to be released from the mitochondria to the cytosol following potent activation of caspase-3 and -9 by typical DNA fragmentation. This apoptosis was regulated by the JNK-signaling pathway. In the present experiment, binding of GSH-DXR to GST P1-1 allosterically led to the disappearance of its enzyme activity and activated the kinase activity of JNK without dissociation of the JNK-GST P1-1 complex. The recombinant GST P1-1 molecule with a mutation in the active center region (W38H and C47S) lost its GST activity when bound to JNK to the same degree as the wild-type, with the mutated GST P1-1 molecule failing to inhibit the activity of JNK. It has been reported that JNK-signaling is regulated by GST P1-1 via interaction with the C-terminus. We confirmed that GST P1-1 deletion mutant (Δ194–209) and a site-directed mutant (R201A) in the C-terminal region failed to bind and inhibit JNK. These results indicated that not only binding of the C-terminal region of GST P1-1 to the JNK molecule, but also the active center region of GST P1-1 play important roles in the regulation of JNK enzyme activity. The findings suggested that allosteric inhibition of GST P1-1 activity by the binding of GSH-DXR following conformational change may activate JNK and induce apoptosis via the mitochondrial pathway in the cells.

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Authors and Affiliations

  1. Department of Biochemistry (I), Jikei University School of Medicine, 3-25-8 Nishi-shinbashi, Minato-ku, Tokyo, 105-8461, Japan

    Tadashi Asakura, Atsuko Sasagawa, Hitoshi Takeuchi, Shun-ichi Shibata, Hideki Marushima, Satoshi Mamori & Kiyoshi Ohkawa

  2. Department of Pharmaceutical Therapeutics, Kyoritsu College of Pharmacy, Tokyo, 105-8512, Japan

    Atsuko Sasagawa & Hitoshi Takeuchi

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  1. Tadashi Asakura
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  2. Atsuko Sasagawa
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Correspondence to Tadashi Asakura.

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Asakura, T., Sasagawa, A., Takeuchi, H. et al. Conformational change in the active center region of GST P1-1, due to binding of a synthetic conjugate of DXR with GSH, enhanced JNK-mediated apoptosis. Apoptosis 12, 1269–1280 (2007). https://doi.org/10.1007/s10495-007-0053-0

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