The structural and biological significance of the EAAEAE insert in the α-domain of human neuronal growth inhibitory factor

Bin Cai, Zhi-Chun Ding, Qi Zhang, Feng Yun Ni, Hui Wang, Qi Zheng, Yang Wang, Guo Ming Zhou, Ke Qiang Wang, Hong Zhe Sun, Hou Ming Wu, Zhong Xian Huang

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Human neuronal growth inhibitory factor (hGIF) is able to inhibit the outgrowth of neurons. As compared with the amino acid sequences of metallothionein 1/2, hGIF contains two insertions: a Thr at position 5 and an acidic hexapeptide EAAEAE(55-60) close to the C-terminus. Moreover, all mammalian growth inhibitory factor sequences contain a conserved CPCP(6-9) motif. Previous studies have demonstrated that the TCPCP(5-9) motif is pivotal to its bioactivity, but no specific role has been assigned to the unique EAAEAE(55-60) insert. To investigate the potential structural and biological significance of the EAAEAE(55-60) insert, several mutants were constructed and investigated in detail. Notably, deletion of the acidic insert (the Δ55-60 mutant) reduced the inhibitory activity, whereas the bioactivities of other mutants did not change much. Then, spectroscopic characterization and molecular dynamics simulation were performed to investigate the potential causes of the reduced bioactivity of the Δ55-60 mutant. It was found that the domain-domain interaction mechanism of hGIF was different from that of metallothionein 2. It was also shown that the acidic insert could regulate the interdomain interactions in hGIF, leading to the structural change in the β-domain, which resulted in the alteration of the solvent accessibility and metal release ability, thus playing an important role in the biological activity of hGIF. Our studies provided useful information on the domain-domain interaction at the molecular level for the first time, and shed new light on the mechanism of the bioactivity of hGIF.

Original languageEnglish (US)
Pages (from-to)3547-3558
Number of pages12
JournalFEBS Journal
Volume276
Issue number13
DOIs
StatePublished - Jan 1 2009

Fingerprint

Bioactivity
Metallothionein
Molecular Dynamics Simulation
growth inhibitory factor
Human Activities
Neurons
Molecular dynamics
Amino Acid Sequence
Metals
Amino Acids
Computer simulation

Keywords

  • Cell culture
  • EAAEAE insert
  • Metallothionein (MT)
  • Molecular dynamics simulation
  • Neuronal growth inhibitory factor (GIF)

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

The structural and biological significance of the EAAEAE insert in the α-domain of human neuronal growth inhibitory factor. / Cai, Bin; Ding, Zhi-Chun; Zhang, Qi; Ni, Feng Yun; Wang, Hui; Zheng, Qi; Wang, Yang; Zhou, Guo Ming; Wang, Ke Qiang; Sun, Hong Zhe; Wu, Hou Ming; Huang, Zhong Xian.

In: FEBS Journal, Vol. 276, No. 13, 01.01.2009, p. 3547-3558.

Research output: Contribution to journalArticle

Cai, B, Ding, Z-C, Zhang, Q, Ni, FY, Wang, H, Zheng, Q, Wang, Y, Zhou, GM, Wang, KQ, Sun, HZ, Wu, HM & Huang, ZX 2009, 'The structural and biological significance of the EAAEAE insert in the α-domain of human neuronal growth inhibitory factor', FEBS Journal, vol. 276, no. 13, pp. 3547-3558. https://doi.org/10.1111/j.1742-4658.2009.07075.x
Cai, Bin ; Ding, Zhi-Chun ; Zhang, Qi ; Ni, Feng Yun ; Wang, Hui ; Zheng, Qi ; Wang, Yang ; Zhou, Guo Ming ; Wang, Ke Qiang ; Sun, Hong Zhe ; Wu, Hou Ming ; Huang, Zhong Xian. / The structural and biological significance of the EAAEAE insert in the α-domain of human neuronal growth inhibitory factor. In: FEBS Journal. 2009 ; Vol. 276, No. 13. pp. 3547-3558.
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AU - Cai, Bin

AU - Ding, Zhi-Chun

AU - Zhang, Qi

AU - Ni, Feng Yun

AU - Wang, Hui

AU - Zheng, Qi

AU - Wang, Yang

AU - Zhou, Guo Ming

AU - Wang, Ke Qiang

AU - Sun, Hong Zhe

AU - Wu, Hou Ming

AU - Huang, Zhong Xian

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N2 - Human neuronal growth inhibitory factor (hGIF) is able to inhibit the outgrowth of neurons. As compared with the amino acid sequences of metallothionein 1/2, hGIF contains two insertions: a Thr at position 5 and an acidic hexapeptide EAAEAE(55-60) close to the C-terminus. Moreover, all mammalian growth inhibitory factor sequences contain a conserved CPCP(6-9) motif. Previous studies have demonstrated that the TCPCP(5-9) motif is pivotal to its bioactivity, but no specific role has been assigned to the unique EAAEAE(55-60) insert. To investigate the potential structural and biological significance of the EAAEAE(55-60) insert, several mutants were constructed and investigated in detail. Notably, deletion of the acidic insert (the Δ55-60 mutant) reduced the inhibitory activity, whereas the bioactivities of other mutants did not change much. Then, spectroscopic characterization and molecular dynamics simulation were performed to investigate the potential causes of the reduced bioactivity of the Δ55-60 mutant. It was found that the domain-domain interaction mechanism of hGIF was different from that of metallothionein 2. It was also shown that the acidic insert could regulate the interdomain interactions in hGIF, leading to the structural change in the β-domain, which resulted in the alteration of the solvent accessibility and metal release ability, thus playing an important role in the biological activity of hGIF. Our studies provided useful information on the domain-domain interaction at the molecular level for the first time, and shed new light on the mechanism of the bioactivity of hGIF.

AB - Human neuronal growth inhibitory factor (hGIF) is able to inhibit the outgrowth of neurons. As compared with the amino acid sequences of metallothionein 1/2, hGIF contains two insertions: a Thr at position 5 and an acidic hexapeptide EAAEAE(55-60) close to the C-terminus. Moreover, all mammalian growth inhibitory factor sequences contain a conserved CPCP(6-9) motif. Previous studies have demonstrated that the TCPCP(5-9) motif is pivotal to its bioactivity, but no specific role has been assigned to the unique EAAEAE(55-60) insert. To investigate the potential structural and biological significance of the EAAEAE(55-60) insert, several mutants were constructed and investigated in detail. Notably, deletion of the acidic insert (the Δ55-60 mutant) reduced the inhibitory activity, whereas the bioactivities of other mutants did not change much. Then, spectroscopic characterization and molecular dynamics simulation were performed to investigate the potential causes of the reduced bioactivity of the Δ55-60 mutant. It was found that the domain-domain interaction mechanism of hGIF was different from that of metallothionein 2. It was also shown that the acidic insert could regulate the interdomain interactions in hGIF, leading to the structural change in the β-domain, which resulted in the alteration of the solvent accessibility and metal release ability, thus playing an important role in the biological activity of hGIF. Our studies provided useful information on the domain-domain interaction at the molecular level for the first time, and shed new light on the mechanism of the bioactivity of hGIF.

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