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Involvement of the residues of GSKIP, AxinGID, and FRATtide in their binding with GSK3β to unravel a novel C-terminal scaffold-binding region

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Abstract

The specificity and regulation of GSK3β are thought to involve in the docking interactions at core kinase domain because of the particular amino acid residues. Recent X-ray diffraction studies illuminated the relative binding residues on AxinGID and FRATtide for GSK3β docking and appeared that GSK3β Val267Gly (V267G) and Tyr288Phe (Y288F) could distinguish the direct interaction between AxinGID and FRATtide. In order to explore the mode that involved the binding of GSKIP to GSK3β and compare it with that of AxinGID and FRATtide, we pinpointed the binding sites of GSKIP to GSK3β through the single-point mutation of four corresponding sites within GSK3β (residues 260–300) as scaffold-binding region I (designated SBR-I260–300). Our data showed that these three binding proteins shared similar binding sites on GSK3β. We also found that the binding of GSK3β V267G mutant to GSKIP and AxinGID, but not that of Y288F mutant (effect on FRATtide), was affected. Further, based on the simulation data, the electron-density map of GSKIPtide bore closer similarity to the map AxinGID than to that of FRATtide. Interestingly, many C-terminal helix region point-mutants of GSK3β L359P, F362A, E366K, and L367P were able to eliminate the binding with FRATtide, but not AxinGID or GSKIP. In addition, CABYR exhibited a unique mode in binding to C-terminal helix region of GSK3β. Taken together, our data revealed that in addition to the core kinase domain, SBR-I260–300, another novel C-terminus helix region, designated SBR-II339–383, also appeared to participate in the recognition and specificity of GSK3β in binding to other specific proteins.

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Acknowledgments

We thank Dr. C. Liu for his helpful comments on the manuscript and Dr. T.C. Dale for providing many of the plasmids of GSK3β mutant listed in Table 1. This study was supported by NSC 95-2314-B-037-050-MY1 (Taiwan, ROC) to SLH, NSC 96-2320-B-037-004, and MOE 97-U19609-1.1ab-14 to YRH.

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Authors and Affiliations

  1. Department of Neurosurgery, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan, ROC

    Shen-Long Howng, Ann-Shung Lieu & Joon-Khim Loh

  2. Department of Biochemistry, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, No. 100, Shih-Chuan 1st Road, Kaohsiung, 80708, Taiwan, ROC

    Chi-Ching Hwang, Chia-Yi Hsu, Meng-Yu Hsu, Chun-Yen Teng, Chia-Hua Chou, Mei-Feng Lee, Chia-Hung Wu, Shean-Jaw Chiou & Yi-Ren Hong

  3. Institute of Biotechnology, National University of Kaohsiung, Kaohsiung, Taiwan, ROC

    Chia-Ning Yang

  4. Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung, Taiwan, ROC

    Chan-Shing Lin

  5. Center of Excellence for Environmental Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan, ROC

    Yi-Ren Hong

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  1. Shen-Long Howng
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Correspondence to Yi-Ren Hong.

Additional information

S.-L. Howng, C.-C. Hwang, and C.-Y. Hsu contributed equally to the work.

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Howng, SL., Hwang, CC., Hsu, CY. et al. Involvement of the residues of GSKIP, AxinGID, and FRATtide in their binding with GSK3β to unravel a novel C-terminal scaffold-binding region. Mol Cell Biochem 339, 23–33 (2010). https://doi.org/10.1007/s11010-009-0366-0

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  • DOI: https://doi.org/10.1007/s11010-009-0366-0

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