Identification of a zinc finger domain in the human NEIL2 (Nei-like-2) protein

Aditi Das, Lavanya Rajagopalan, Venkatarajan S. Mathura, Samuel J. Rigby, Sankar Mitra, Tapas Hazra

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

The recently identified human NEIL2 (Nei-like-2) protein, a DNA glycosylase/AP lyase specific for oxidatively damaged bases, shares structural features and reaction mechanism with the Escherichia coli DNA glycosylases, Nei and Fpg. Amino acid sequence analysis of NEIL2 suggested it to have a zinc finger-like Nei/Fpg. However, the Cys-X2-His-X16-Cys- X2-Cys (CHCC) motif present near the C terminus of NEIL2 is distinct from the zinc finger motifs of Nei/Fpg, which are of the C4 type. Here we show the presence of an equimolar amount of zinc in NEIL2 by inductively coupled plasma mass spectrometry. Individual mutations of Cys-291, His-295, Cys-315, and Cys-318, candidate residues for coordinating zinc, inactivated the enzyme by abolishing its DNA binding activity. H295A and C318S mutants were also shown to lack bound zinc, and a significant change in their secondary structure was revealed by CD spectra analysis. Molecular modeling revealed Arg-310 of NEIL2 to be a critical residue in its zinc binding pocket, which is highly conserved throughout the Fpg/Nei family. A R310Q mutation significantly reduced the activity of NEIL2. We thereby conclude that the zinc finger motif in NEIL2 is essential for its structural integrity and enzyme activity.

Original languageEnglish (US)
Pages (from-to)47132-47138
Number of pages7
JournalJournal of Biological Chemistry
Volume279
Issue number45
DOIs
StatePublished - Nov 5 2004

Fingerprint

Zinc Fingers
Zinc
DNA Glycosylases
DNA-(Apurinic or Apyrimidinic Site) Lyase
Mutation
Protein Sequence Analysis
Enzymes
Mass Spectrometry
Spectrum Analysis
Inductively coupled plasma mass spectrometry
Escherichia coli
Molecular modeling
Enzyme activity
Structural integrity
human NEIL2 protein
DNA
Spectrum analysis
Amino Acids

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification of a zinc finger domain in the human NEIL2 (Nei-like-2) protein. / Das, Aditi; Rajagopalan, Lavanya; Mathura, Venkatarajan S.; Rigby, Samuel J.; Mitra, Sankar; Hazra, Tapas.

In: Journal of Biological Chemistry, Vol. 279, No. 45, 05.11.2004, p. 47132-47138.

Research output: Contribution to journalArticle

Das, Aditi ; Rajagopalan, Lavanya ; Mathura, Venkatarajan S. ; Rigby, Samuel J. ; Mitra, Sankar ; Hazra, Tapas. / Identification of a zinc finger domain in the human NEIL2 (Nei-like-2) protein. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 45. pp. 47132-47138.
@article{ee5654b404dd45f59aaeca55490e7cc0,
title = "Identification of a zinc finger domain in the human NEIL2 (Nei-like-2) protein",
abstract = "The recently identified human NEIL2 (Nei-like-2) protein, a DNA glycosylase/AP lyase specific for oxidatively damaged bases, shares structural features and reaction mechanism with the Escherichia coli DNA glycosylases, Nei and Fpg. Amino acid sequence analysis of NEIL2 suggested it to have a zinc finger-like Nei/Fpg. However, the Cys-X2-His-X16-Cys- X2-Cys (CHCC) motif present near the C terminus of NEIL2 is distinct from the zinc finger motifs of Nei/Fpg, which are of the C4 type. Here we show the presence of an equimolar amount of zinc in NEIL2 by inductively coupled plasma mass spectrometry. Individual mutations of Cys-291, His-295, Cys-315, and Cys-318, candidate residues for coordinating zinc, inactivated the enzyme by abolishing its DNA binding activity. H295A and C318S mutants were also shown to lack bound zinc, and a significant change in their secondary structure was revealed by CD spectra analysis. Molecular modeling revealed Arg-310 of NEIL2 to be a critical residue in its zinc binding pocket, which is highly conserved throughout the Fpg/Nei family. A R310Q mutation significantly reduced the activity of NEIL2. We thereby conclude that the zinc finger motif in NEIL2 is essential for its structural integrity and enzyme activity.",
author = "Aditi Das and Lavanya Rajagopalan and Mathura, {Venkatarajan S.} and Rigby, {Samuel J.} and Sankar Mitra and Tapas Hazra",
year = "2004",
month = "11",
day = "5",
doi = "10.1074/jbc.M406224200",
language = "English (US)",
volume = "279",
pages = "47132--47138",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "45",

}

TY - JOUR

T1 - Identification of a zinc finger domain in the human NEIL2 (Nei-like-2) protein

AU - Das, Aditi

AU - Rajagopalan, Lavanya

AU - Mathura, Venkatarajan S.

AU - Rigby, Samuel J.

AU - Mitra, Sankar

AU - Hazra, Tapas

PY - 2004/11/5

Y1 - 2004/11/5

N2 - The recently identified human NEIL2 (Nei-like-2) protein, a DNA glycosylase/AP lyase specific for oxidatively damaged bases, shares structural features and reaction mechanism with the Escherichia coli DNA glycosylases, Nei and Fpg. Amino acid sequence analysis of NEIL2 suggested it to have a zinc finger-like Nei/Fpg. However, the Cys-X2-His-X16-Cys- X2-Cys (CHCC) motif present near the C terminus of NEIL2 is distinct from the zinc finger motifs of Nei/Fpg, which are of the C4 type. Here we show the presence of an equimolar amount of zinc in NEIL2 by inductively coupled plasma mass spectrometry. Individual mutations of Cys-291, His-295, Cys-315, and Cys-318, candidate residues for coordinating zinc, inactivated the enzyme by abolishing its DNA binding activity. H295A and C318S mutants were also shown to lack bound zinc, and a significant change in their secondary structure was revealed by CD spectra analysis. Molecular modeling revealed Arg-310 of NEIL2 to be a critical residue in its zinc binding pocket, which is highly conserved throughout the Fpg/Nei family. A R310Q mutation significantly reduced the activity of NEIL2. We thereby conclude that the zinc finger motif in NEIL2 is essential for its structural integrity and enzyme activity.

AB - The recently identified human NEIL2 (Nei-like-2) protein, a DNA glycosylase/AP lyase specific for oxidatively damaged bases, shares structural features and reaction mechanism with the Escherichia coli DNA glycosylases, Nei and Fpg. Amino acid sequence analysis of NEIL2 suggested it to have a zinc finger-like Nei/Fpg. However, the Cys-X2-His-X16-Cys- X2-Cys (CHCC) motif present near the C terminus of NEIL2 is distinct from the zinc finger motifs of Nei/Fpg, which are of the C4 type. Here we show the presence of an equimolar amount of zinc in NEIL2 by inductively coupled plasma mass spectrometry. Individual mutations of Cys-291, His-295, Cys-315, and Cys-318, candidate residues for coordinating zinc, inactivated the enzyme by abolishing its DNA binding activity. H295A and C318S mutants were also shown to lack bound zinc, and a significant change in their secondary structure was revealed by CD spectra analysis. Molecular modeling revealed Arg-310 of NEIL2 to be a critical residue in its zinc binding pocket, which is highly conserved throughout the Fpg/Nei family. A R310Q mutation significantly reduced the activity of NEIL2. We thereby conclude that the zinc finger motif in NEIL2 is essential for its structural integrity and enzyme activity.

UR - http://www.scopus.com/inward/record.url?scp=8744262953&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=8744262953&partnerID=8YFLogxK

U2 - 10.1074/jbc.M406224200

DO - 10.1074/jbc.M406224200

M3 - Article

VL - 279

SP - 47132

EP - 47138

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 45

ER -