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The role of Thr5 in human neuron growth inhibitory factor

JBIC Journal of Biological Inorganic Chemistry volume 11, Article number: 476 (2006) Cite this article

Abstract

GIF, a member of the metallothionein (MT) family (assigned as MT3), is a neuron growth inhibitory factor that inhibits neuron outgrowth in Alzheimer’s disease. The conserved Thr5 is one of the main differences between GIF and other members in the MT family. However, natural sheep GIF has an unusual Ala5, casting doubt on the role of common Thr5. We constructed a series of human GIF mutants at site 5, and characterized their biochemical properties by UV spectroscopy, circular dichroism spectroscopy, EDTA reaction, 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB) reaction, and pH titration. Their inhibitory activity toward neuron survival and neurite extension was also examined. Interestingly, the T5A mutant exhibited distinct metal thiolate activity in the EDTA and DTNB reactions, and also lost its bioactivity. Meanwhile, the T5S mutant had similar biochemical properties and biological activity as wild-type human GIF, indicating the hydroxyl group on the Thr5 was critical to the bioactivity of human GIF. We suggest the hydroxyl group in human GIF may help stabilize the biologically active conformation. On the other hand, lack of the hydroxyl group in sheep GIF may be partially compensated by its abnormal structure.

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Affiliations

  1. Chemical Biology Laboratory, Department of Chemistry, Fudan University, 200433, Shanghai, China

    Bin Cai, Qi Zheng, Xin-Chen Teng, Dong Chen & Zhong-Xian Huang

  2. Institute of Genetics, Life School, Fudan University, 200433, Shanghai, China

    Yi Xie

  3. Department of Anatomy, Histology and Embryology, Medical School, Fudan University, 200032, Shanghai, China

    Yang Wang & Guo-Ming Zhou

  4. Research Institute of Cardiovascular Disease, Zhongshan Hospital, Medical School, Fudan University, 200032, Shanghai, China

    Ke-Qiang Wang

  5. Department of Chemistry, The University of Hong Kong, Pokfulam Road, Hong Kong, China

    Hong-Zhe Sun

  6. Department of Biochemistry, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China

    Ming-Jie Zhang

Authors
  1. Bin Cai
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  2. Qi Zheng
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  3. Xin-Chen Teng
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  4. Dong Chen
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  5. Yang Wang
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  6. Ke-Qiang Wang
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  7. Guo-Ming Zhou
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  8. Yi Xie
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  9. Ming-Jie Zhang
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  10. Hong-Zhe Sun
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  11. Zhong-Xian Huang
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Corresponding authors

Correspondence to Hong-Zhe Sun or Zhong-Xian Huang.

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Bin Cai and Qi Zheng contributed equally to this work.

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Cai, B., Zheng, Q., Teng, XC. et al. The role of Thr5 in human neuron growth inhibitory factor. J Biol Inorg Chem 11, 476 (2006). https://doi.org/10.1007/s00775-006-0097-6

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  • DOI: https://doi.org/10.1007/s00775-006-0097-6

Keywords

  • Neurite Length
  • Neurite Extension
  • Dithiodipyridine
  • Fusion Expression Vector
  • Thiolate Cluster