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Targeting rho guanine nucleotide exchange factor ARHGEF5/TIM with auto-inhibitory peptides in human breast cancer

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Abstract

The oncogenic protein ARHGEF5/TIM has long been known to express specifically in human breast cancer and other tumors, which is an important member of Rho guanine nucleotide exchange factors that activate Rho-family GTPases by promoting GTP/GDP exchange. The activation capability of TIM is auto-inhibited by a putative helix N-terminal to Dbl homology (DH) domain, which is stabilized by intramolecular interaction of Src homology 3 domain with a poly-proline sequence that locates between the helix and DH domain. Here, we attempted to target TIM DH domain using the modified versions of its auto-inhibitory helix. In the procedure, bioinformatics techniques were used to investigate the intramolecular interaction of DH domain with auto-inhibitory helix and, based on obtained knowledge, to optimize physicochemical property and structural conformation for the helix. We also performed affinity assay to determine the binding strength of modified peptides to DH domain. Consequently, two modified peptides, namely, DALYEEYNLVV and EVLYEEYQLVV were found as good binders of DH domain with dissociation constants K d of 0.35 and 2 µM, respectively. Structural analysis revealed that the charge neutralization and electrostatic interaction confer additional stability for these two peptide complexes with DH domain.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (Nos. 30801118, 81172520, 81202087, 81202088, 81472462 and 81402176), the Natural Science Foundation of Shanghai Municipal Science and Technology Commission (No. 12ZR1446400), the Technology Innovation Act Plan of Shanghai Municipal Science and Technology Commission (Nos. 14411950200 and 14411950201), the Wenzhou Science and Technology Bureau (No. Y20130090), the Natural Science Foundation of Jiangsu Province (No. BK20140288), the Natural Science Foundation of Jiangsu Higher Education Institutions of China (No. 14KJB320011), and the Joint Research Project of the Emerging Cutting-edge Technology of Shanghai Shen-kang Hospital Development Center (No. SHDC12014103).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Surgery, Ruijin Hospital, Medical School of Shanghai Jiaotong University, Shanghai, 200025, China

    Ou Huang, Yafen Li, Weiguo Chen & Kunwei Shen

  2. International Peace Maternity and Child Health Hospital, Shanghai Jiaotong University, Shanghai, 200023, China

    Dandan Wu

  3. Department of Surgical Oncology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, 325000, China

    Feiyan Xie, Lili Lin, Xiaobo Wang & Xiaoqu Hu

  4. Department of Breast Surgery, The First Affiliated Hospital of Soochow University, Suzhou, 215006, China

    Min Jiang

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  1. Ou Huang
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  2. Dandan Wu
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  3. Feiyan Xie
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  4. Lili Lin
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  5. Xiaobo Wang
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  6. Min Jiang
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  7. Yafen Li
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  8. Weiguo Chen
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  10. Xiaoqu Hu
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Correspondence to Xiaoqu Hu.

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Handling Editor: M. S. Palma.

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Huang, O., Wu, D., Xie, F. et al. Targeting rho guanine nucleotide exchange factor ARHGEF5/TIM with auto-inhibitory peptides in human breast cancer. Amino Acids 47, 1239–1246 (2015). https://doi.org/10.1007/s00726-015-1950-0

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  • DOI: https://doi.org/10.1007/s00726-015-1950-0

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