Selection of novel structural zinc sites from a random peptide library

Teruhiko Matsubara, Yuko Hiura, Osamu Kawahito, Mikito Yasuzawa, Katsuhiro Kawashiro

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Zinc ion (Zn2+) can be coordinated with four or three amino acid residues to stabilize a protein's structure or to form a catalytic active center. We used phage display selection of a dodecamer random peptide library with Zn2+ to identify structural zinc sites. The binding specificity for Zn2+ of selected sequences was confirmed using enzyme-linked immunosorbent and competitive inhibition assays. Circular dichroism spectra indicated that the interaction with Zn2+ induced a change in conformation, which means the peptide acts as a structural zinc site. Furthermore, a search of protein databases revealed that two selected sequences corresponded to parts of natural zinc sites of copper/zinc superoxide dismutase and zinc-containing ferredoxin. We demonstrated that Zn2+-binding sequences selected from the random combinatorial library would be candidates for artificial structural zinc sites.

Original languageEnglish
Pages (from-to)317-321
Number of pages5
JournalFEBS Letters
Volume555
Issue number2
DOIs
Publication statusPublished - 2003 Dec 4

Fingerprint

Peptide Library
Zinc
Ferredoxins
Immunosorbents
Protein Databases
Bacteriophages
Circular Dichroism
Superoxide Dismutase
Conformations
Copper
Assays
Proteins
Display devices
Ions
Amino Acids
Peptides
Enzymes

Keywords

  • Metalloenzyme
  • Phage display system
  • Random peptide library
  • Sequence homology
  • Structural zinc

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Selection of novel structural zinc sites from a random peptide library. / Matsubara, Teruhiko; Hiura, Yuko; Kawahito, Osamu; Yasuzawa, Mikito; Kawashiro, Katsuhiro.

In: FEBS Letters, Vol. 555, No. 2, 04.12.2003, p. 317-321.

Research output: Contribution to journalArticle

Matsubara, T, Hiura, Y, Kawahito, O, Yasuzawa, M & Kawashiro, K 2003, 'Selection of novel structural zinc sites from a random peptide library', FEBS Letters, vol. 555, no. 2, pp. 317-321. https://doi.org/10.1016/S0014-5793(03)01266-3
Matsubara, Teruhiko ; Hiura, Yuko ; Kawahito, Osamu ; Yasuzawa, Mikito ; Kawashiro, Katsuhiro. / Selection of novel structural zinc sites from a random peptide library. In: FEBS Letters. 2003 ; Vol. 555, No. 2. pp. 317-321.
@article{3af601de494b41ad82f00817ed3a1839,
title = "Selection of novel structural zinc sites from a random peptide library",
abstract = "Zinc ion (Zn2+) can be coordinated with four or three amino acid residues to stabilize a protein's structure or to form a catalytic active center. We used phage display selection of a dodecamer random peptide library with Zn2+ to identify structural zinc sites. The binding specificity for Zn2+ of selected sequences was confirmed using enzyme-linked immunosorbent and competitive inhibition assays. Circular dichroism spectra indicated that the interaction with Zn2+ induced a change in conformation, which means the peptide acts as a structural zinc site. Furthermore, a search of protein databases revealed that two selected sequences corresponded to parts of natural zinc sites of copper/zinc superoxide dismutase and zinc-containing ferredoxin. We demonstrated that Zn2+-binding sequences selected from the random combinatorial library would be candidates for artificial structural zinc sites.",
keywords = "Metalloenzyme, Phage display system, Random peptide library, Sequence homology, Structural zinc",
author = "Teruhiko Matsubara and Yuko Hiura and Osamu Kawahito and Mikito Yasuzawa and Katsuhiro Kawashiro",
year = "2003",
month = "12",
day = "4",
doi = "10.1016/S0014-5793(03)01266-3",
language = "English",
volume = "555",
pages = "317--321",
journal = "FEBS Letters",
issn = "0014-5793",
publisher = "Elsevier",
number = "2",

}

TY - JOUR

T1 - Selection of novel structural zinc sites from a random peptide library

AU - Matsubara, Teruhiko

AU - Hiura, Yuko

AU - Kawahito, Osamu

AU - Yasuzawa, Mikito

AU - Kawashiro, Katsuhiro

PY - 2003/12/4

Y1 - 2003/12/4

N2 - Zinc ion (Zn2+) can be coordinated with four or three amino acid residues to stabilize a protein's structure or to form a catalytic active center. We used phage display selection of a dodecamer random peptide library with Zn2+ to identify structural zinc sites. The binding specificity for Zn2+ of selected sequences was confirmed using enzyme-linked immunosorbent and competitive inhibition assays. Circular dichroism spectra indicated that the interaction with Zn2+ induced a change in conformation, which means the peptide acts as a structural zinc site. Furthermore, a search of protein databases revealed that two selected sequences corresponded to parts of natural zinc sites of copper/zinc superoxide dismutase and zinc-containing ferredoxin. We demonstrated that Zn2+-binding sequences selected from the random combinatorial library would be candidates for artificial structural zinc sites.

AB - Zinc ion (Zn2+) can be coordinated with four or three amino acid residues to stabilize a protein's structure or to form a catalytic active center. We used phage display selection of a dodecamer random peptide library with Zn2+ to identify structural zinc sites. The binding specificity for Zn2+ of selected sequences was confirmed using enzyme-linked immunosorbent and competitive inhibition assays. Circular dichroism spectra indicated that the interaction with Zn2+ induced a change in conformation, which means the peptide acts as a structural zinc site. Furthermore, a search of protein databases revealed that two selected sequences corresponded to parts of natural zinc sites of copper/zinc superoxide dismutase and zinc-containing ferredoxin. We demonstrated that Zn2+-binding sequences selected from the random combinatorial library would be candidates for artificial structural zinc sites.

KW - Metalloenzyme

KW - Phage display system

KW - Random peptide library

KW - Sequence homology

KW - Structural zinc

UR - http://www.scopus.com/inward/record.url?scp=0344009452&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0344009452&partnerID=8YFLogxK

U2 - 10.1016/S0014-5793(03)01266-3

DO - 10.1016/S0014-5793(03)01266-3

M3 - Article

C2 - 14644435

AN - SCOPUS:0344009452

VL - 555

SP - 317

EP - 321

JO - FEBS Letters

JF - FEBS Letters

SN - 0014-5793

IS - 2

ER -