Incorporation of a glycine within the conserved TCPCP motif of human neuronal growth inhibitory factor significantly reduces its bioactivity

Zhi-Chun Ding, Dong Chen, Feng Yun Ni, Qi Zheng, Bin Cai, Wen Hua Yao, Yang Wang, Guo Ming Zhou, Zhong Xian Huang

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

It has been reported that the 6 CPCP 9 motif near the N-terminus is pivotal to the inhibitory activity of human neuronal growth inhibitory factor (hGIF). In order to better understand the biological significance of this region on the structure, property and function of hGIF, we introduced a highly flexible residue, Gly, either in front of the 6 CPCP 9 motif (the IG6 mutant, TGCPCP) or in the middle of it (the IG8 mutant, TCPGCP) and investigated their structural and metal binding properties in detail. The results showed that the overall structure and the stability of the metal-thiolate clusters of the two mutants were comparable to that of hGIF. However, the bioassay results showed that the bioactivity of the IG6 mutant decreased significantly, while the bioactivity of the IG8 mutant was almost abolished. Molecular dynamics simulation results showed that the backbone of the IG6 mutant exhibited high similarity to that of hGIF, and the two prolines could still induce structural constraints on the 6 CPCP 9 tetrapeptide and form a similar conformation with that of hGIF, however, the conformation of the first five amino acid residues in the N-terminus was quite different. In hGIF, the five residues are twisted and form a restricted conformation, while in the IG6 mutant this peptide extends more naturally and smoothly, which is similar to that of MT2. As to the IG8 mutant, the Gly insertion broke the 6 CPCP 9 motif, thus probably abolishing the interactions with other molecules and eliminating its inhibitory activity. Based on these results, we suggested that although the structure adopted by the 6 CPCP 9 motif is the determinant factor of the inhibitory bioactivity of hGIF, other residues within the N-terminal fragment (residue 1-13) may also influence the peptide conformation and contribute to the protein's bioactivity.

Original languageEnglish (US)
Pages (from-to)779-784
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume372
Issue number4
DOIs
StatePublished - Aug 8 2008

Fingerprint

Bioactivity
Glycine
Conformations
Metals
Peptides
Bioassay
Molecular Dynamics Simulation
growth inhibitory factor
Proline
Human Activities
Biological Assay
Molecular dynamics
Amino Acids
Molecules
Computer simulation
Proteins

Keywords

  • Metallothionein
  • Molecular dynamics simulation
  • Mutants
  • Neuronal cell culture
  • Neuronal growth inhibitory factor (GIF)

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Incorporation of a glycine within the conserved TCPCP motif of human neuronal growth inhibitory factor significantly reduces its bioactivity. / Ding, Zhi-Chun; Chen, Dong; Ni, Feng Yun; Zheng, Qi; Cai, Bin; Yao, Wen Hua; Wang, Yang; Zhou, Guo Ming; Huang, Zhong Xian.

In: Biochemical and Biophysical Research Communications, Vol. 372, No. 4, 08.08.2008, p. 779-784.

Research output: Contribution to journalArticle

Ding, Zhi-Chun ; Chen, Dong ; Ni, Feng Yun ; Zheng, Qi ; Cai, Bin ; Yao, Wen Hua ; Wang, Yang ; Zhou, Guo Ming ; Huang, Zhong Xian. / Incorporation of a glycine within the conserved TCPCP motif of human neuronal growth inhibitory factor significantly reduces its bioactivity. In: Biochemical and Biophysical Research Communications. 2008 ; Vol. 372, No. 4. pp. 779-784.
@article{6bc5289db7fa426f9bd77e1e9953104e,
title = "Incorporation of a glycine within the conserved TCPCP motif of human neuronal growth inhibitory factor significantly reduces its bioactivity",
abstract = "It has been reported that the 6 CPCP 9 motif near the N-terminus is pivotal to the inhibitory activity of human neuronal growth inhibitory factor (hGIF). In order to better understand the biological significance of this region on the structure, property and function of hGIF, we introduced a highly flexible residue, Gly, either in front of the 6 CPCP 9 motif (the IG6 mutant, TGCPCP) or in the middle of it (the IG8 mutant, TCPGCP) and investigated their structural and metal binding properties in detail. The results showed that the overall structure and the stability of the metal-thiolate clusters of the two mutants were comparable to that of hGIF. However, the bioassay results showed that the bioactivity of the IG6 mutant decreased significantly, while the bioactivity of the IG8 mutant was almost abolished. Molecular dynamics simulation results showed that the backbone of the IG6 mutant exhibited high similarity to that of hGIF, and the two prolines could still induce structural constraints on the 6 CPCP 9 tetrapeptide and form a similar conformation with that of hGIF, however, the conformation of the first five amino acid residues in the N-terminus was quite different. In hGIF, the five residues are twisted and form a restricted conformation, while in the IG6 mutant this peptide extends more naturally and smoothly, which is similar to that of MT2. As to the IG8 mutant, the Gly insertion broke the 6 CPCP 9 motif, thus probably abolishing the interactions with other molecules and eliminating its inhibitory activity. Based on these results, we suggested that although the structure adopted by the 6 CPCP 9 motif is the determinant factor of the inhibitory bioactivity of hGIF, other residues within the N-terminal fragment (residue 1-13) may also influence the peptide conformation and contribute to the protein's bioactivity.",
keywords = "Metallothionein, Molecular dynamics simulation, Mutants, Neuronal cell culture, Neuronal growth inhibitory factor (GIF)",
author = "Zhi-Chun Ding and Dong Chen and Ni, {Feng Yun} and Qi Zheng and Bin Cai and Yao, {Wen Hua} and Yang Wang and Zhou, {Guo Ming} and Huang, {Zhong Xian}",
year = "2008",
month = "8",
day = "8",
doi = "10.1016/j.bbrc.2008.05.136",
language = "English (US)",
volume = "372",
pages = "779--784",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "4",

}

TY - JOUR

T1 - Incorporation of a glycine within the conserved TCPCP motif of human neuronal growth inhibitory factor significantly reduces its bioactivity

AU - Ding, Zhi-Chun

AU - Chen, Dong

AU - Ni, Feng Yun

AU - Zheng, Qi

AU - Cai, Bin

AU - Yao, Wen Hua

AU - Wang, Yang

AU - Zhou, Guo Ming

AU - Huang, Zhong Xian

PY - 2008/8/8

Y1 - 2008/8/8

N2 - It has been reported that the 6 CPCP 9 motif near the N-terminus is pivotal to the inhibitory activity of human neuronal growth inhibitory factor (hGIF). In order to better understand the biological significance of this region on the structure, property and function of hGIF, we introduced a highly flexible residue, Gly, either in front of the 6 CPCP 9 motif (the IG6 mutant, TGCPCP) or in the middle of it (the IG8 mutant, TCPGCP) and investigated their structural and metal binding properties in detail. The results showed that the overall structure and the stability of the metal-thiolate clusters of the two mutants were comparable to that of hGIF. However, the bioassay results showed that the bioactivity of the IG6 mutant decreased significantly, while the bioactivity of the IG8 mutant was almost abolished. Molecular dynamics simulation results showed that the backbone of the IG6 mutant exhibited high similarity to that of hGIF, and the two prolines could still induce structural constraints on the 6 CPCP 9 tetrapeptide and form a similar conformation with that of hGIF, however, the conformation of the first five amino acid residues in the N-terminus was quite different. In hGIF, the five residues are twisted and form a restricted conformation, while in the IG6 mutant this peptide extends more naturally and smoothly, which is similar to that of MT2. As to the IG8 mutant, the Gly insertion broke the 6 CPCP 9 motif, thus probably abolishing the interactions with other molecules and eliminating its inhibitory activity. Based on these results, we suggested that although the structure adopted by the 6 CPCP 9 motif is the determinant factor of the inhibitory bioactivity of hGIF, other residues within the N-terminal fragment (residue 1-13) may also influence the peptide conformation and contribute to the protein's bioactivity.

AB - It has been reported that the 6 CPCP 9 motif near the N-terminus is pivotal to the inhibitory activity of human neuronal growth inhibitory factor (hGIF). In order to better understand the biological significance of this region on the structure, property and function of hGIF, we introduced a highly flexible residue, Gly, either in front of the 6 CPCP 9 motif (the IG6 mutant, TGCPCP) or in the middle of it (the IG8 mutant, TCPGCP) and investigated their structural and metal binding properties in detail. The results showed that the overall structure and the stability of the metal-thiolate clusters of the two mutants were comparable to that of hGIF. However, the bioassay results showed that the bioactivity of the IG6 mutant decreased significantly, while the bioactivity of the IG8 mutant was almost abolished. Molecular dynamics simulation results showed that the backbone of the IG6 mutant exhibited high similarity to that of hGIF, and the two prolines could still induce structural constraints on the 6 CPCP 9 tetrapeptide and form a similar conformation with that of hGIF, however, the conformation of the first five amino acid residues in the N-terminus was quite different. In hGIF, the five residues are twisted and form a restricted conformation, while in the IG6 mutant this peptide extends more naturally and smoothly, which is similar to that of MT2. As to the IG8 mutant, the Gly insertion broke the 6 CPCP 9 motif, thus probably abolishing the interactions with other molecules and eliminating its inhibitory activity. Based on these results, we suggested that although the structure adopted by the 6 CPCP 9 motif is the determinant factor of the inhibitory bioactivity of hGIF, other residues within the N-terminal fragment (residue 1-13) may also influence the peptide conformation and contribute to the protein's bioactivity.

KW - Metallothionein

KW - Molecular dynamics simulation

KW - Mutants

KW - Neuronal cell culture

KW - Neuronal growth inhibitory factor (GIF)

UR - http://www.scopus.com/inward/record.url?scp=45449083790&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=45449083790&partnerID=8YFLogxK

U2 - 10.1016/j.bbrc.2008.05.136

DO - 10.1016/j.bbrc.2008.05.136

M3 - Article

C2 - 18533104

AN - SCOPUS:45449083790

VL - 372

SP - 779

EP - 784

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 4

ER -