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Inter-molecular crosslinking activity is engendered by the dimeric form of transglutaminase 2

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Abstract

Transglutaminase 2 (TGase 2) catalyzes a crosslink between protein bound-glutamine and -lysine. We proposed the mechanism of TGase 2 activation depends on conformation change from unfolded monomer to unfolded dimer. We found that TGase 2 has temperature-sensitive conformation change system at 30 °C. Small-angle X-ray scattering analysis showed that the enzyme was maintained as an unfolded monomer at temperatures below 30 °C, but changed to an unfolded dimer at over 30 °C. Mass analysis revealed that the C-terminus of TGase 2 was the critical region for dimerization. Furthermore, this conformational switch creates new biochemical reactivity that catalyzed inter-molecular crosslink at above 30 °C as an unfolded dimer of TGase 2 while catalyzed intra-molecular crosslink at below 30 °C as an unfolded monomer of TGase 2. The mechanism of TGase 2 activation depends on temperature-sensitive conformation change from unfolded monomer to unfolded dimer at over 30 °C. Furthermore, inter-molecular crosslinking activity is generated by the dimeric form of TGase 2. TGase 2 switches its conformation from a monomer to a dimer following a change in temperature, which engendered unique catalytic function of enzyme as inter-molecular crosslinking activity with calcium.

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Acknowledgments

The authors thank the staff at Beamline 4C of the Pohang Light Source for their assistance during the SAXS experiments. This research was supported by a grant from the National Cancer Center of Korea to S.-Y.K. (NCC1410280-3).

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    Authors and Affiliations

    1. Cancer Cell and Molecular Biology Branch, Division of Cancer Biology, Research Institute, National Cancer Center, Goyang, 10408, Republic of Korea

      Nayeon Kim, Won-Kyu Lee, Seon-Hyeong Lee & Soo-Youl Kim

    2. Department of Chemistry, College of Science, Dongguk University, Seoul, 04620, Republic of Korea

      Nayeon Kim

    3. Institute of Life Science and Natural Resources, Korea University, Seoul, 02841, Republic of Korea

      Won-Kyu Lee

    4. Pohang Accelerator Laboratory, Pohang University of Science and Technology, 80 Jigokro-127-beongil, Pohang, 37673, Republic of Korea

      Kyeong Sik Jin

    5. Omics Core Laboratory, Research Institute, National Cancer Center, Goyang, 10408, Republic of Korea

      Kyung-Hee Kim

    6. New Drug Development Center, Daegu-Gyeongbuk Medical Innovation Foundation, 80 Cheombok-ro, Dong-gu, Daegu, 41061, Republic of Korea

      Younho Lee & Minsoo Song

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    1. Nayeon Kim
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    Correspondence to Soo-Youl Kim.

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    Handling Editors: S. Beninati, M. Piacentini and C.M. Bergamini.

    N. Kim and W.-K. Lee contributed equally to this study.

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    Kim, N., Lee, WK., Lee, SH. et al. Inter-molecular crosslinking activity is engendered by the dimeric form of transglutaminase 2. Amino Acids 49, 461–471 (2017). https://doi.org/10.1007/s00726-016-2293-1

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