Important roles of the conserved linker-KKS in human neuronal growth inhibitory factor

Zhi-Chun Ding, Xin Chen Teng, Qi Zheng, Feng Yun Ni, Bin Cai, Yang Wang, Guo Ming Zhou, Hong Zhe Sun, Xiang Shi Tan, Zhong Xian Huang

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Metallothinein-3 (MT3), also named neuronal growth inhibitory factor (GIF), is attractive by its distinct neuronal growth inhibitory activity, which is not shared by other MT isoforms. The polypeptide chain of GIF is folded into two individual domains, which are connected by a highly conserved linker, KKS. In order to figure out the significance of the conserved segment, we constructed several mutants of human GIF (hGIF), including the K31/32A mutant, the K31/32E mutant and the KKS-SP mutant by site-directed mutagenesis. pH titration and DTNB reaction exhibited that all the three mutations made the β-domain lower in stability and looser. More significantly, change of KKS to SP also altered the general backbone conformation and metal-thiolate cluster geometry. Notably, bioassay results showed that the bioactivity of the K31/32A mutant and the K31/32E mutant decreased obviously, while the KKS-SP mutant lost inhibitory activity completely. Based on these results, we proposed that the KKS linker was a crucial factor in modulating the stability and the solvent accessibility of the Cd 3 S 9 cluster in the β-domain through domain-domain interactions, thus was indispensable to the biological activity of hGIF.

Original languageEnglish (US)
Pages (from-to)817-826
Number of pages10
JournalBioMetals
Volume22
Issue number5
DOIs
StatePublished - Oct 1 2009

Fingerprint

Bioactivity
mutants
Mutagenesis
Bioassay
Polypeptides
Dithionitrobenzoic Acid
Titration
Conformations
Site-Directed Mutagenesis
Biological Assay
Protein Isoforms
Geometry
Metals
human growth
site-directed mutagenesis
Peptides
Mutation
titration
Intercellular Signaling Peptides and Proteins
growth inhibitory factor

Keywords

  • Cell culture
  • Linker
  • Metallothionein
  • Mutation
  • Neuronal growth inhibitory factor

ASJC Scopus subject areas

  • Biomaterials
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • Metals and Alloys

Cite this

Ding, Z-C., Teng, X. C., Zheng, Q., Ni, F. Y., Cai, B., Wang, Y., ... Huang, Z. X. (2009). Important roles of the conserved linker-KKS in human neuronal growth inhibitory factor. BioMetals, 22(5), 817-826. https://doi.org/10.1007/s10534-009-9228-1

Important roles of the conserved linker-KKS in human neuronal growth inhibitory factor. / Ding, Zhi-Chun; Teng, Xin Chen; Zheng, Qi; Ni, Feng Yun; Cai, Bin; Wang, Yang; Zhou, Guo Ming; Sun, Hong Zhe; Tan, Xiang Shi; Huang, Zhong Xian.

In: BioMetals, Vol. 22, No. 5, 01.10.2009, p. 817-826.

Research output: Contribution to journalArticle

Ding, Z-C, Teng, XC, Zheng, Q, Ni, FY, Cai, B, Wang, Y, Zhou, GM, Sun, HZ, Tan, XS & Huang, ZX 2009, 'Important roles of the conserved linker-KKS in human neuronal growth inhibitory factor', BioMetals, vol. 22, no. 5, pp. 817-826. https://doi.org/10.1007/s10534-009-9228-1
Ding, Zhi-Chun ; Teng, Xin Chen ; Zheng, Qi ; Ni, Feng Yun ; Cai, Bin ; Wang, Yang ; Zhou, Guo Ming ; Sun, Hong Zhe ; Tan, Xiang Shi ; Huang, Zhong Xian. / Important roles of the conserved linker-KKS in human neuronal growth inhibitory factor. In: BioMetals. 2009 ; Vol. 22, No. 5. pp. 817-826.
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