Biochemical and physicochemical characterization of normal and variant forms of human MTH1 protein with antimutagenic activity

Hiroyuki Yakushiji, Fabrice Maraboeuf, Masayuki Takahashi, Zeng Sui Deng, Shun-Ichiro Kawabata, Yusaku Nakabeppu, Mutsuo Sekiguchi

研究成果: ジャーナルへの寄稿記事

43 引用 (Scopus)

抄録

8-0xo-7,8-dihydro-2'-deoxyguanosine 5'-triphosphate (8-oxo-dGTP) is produced during cellular metabolism, and its misincorporation into DNA causes mutation. Human cells possess an enzyme that hydrolyzes 8-oxo-dGTP to the corresponding nucleoside monophosphate, thereby preventing misincorporation of 8-oxo-7,8-dihydroguanine into DNA. Sequence an analyses of the MTH1 gene, encoding the 8-oxo-7,8-dihydro-2'-deoxyguanosine 5'-triphosphatase (8-oxo-dGTPase) protein in human cell lines revealed that a G to A base substitution frequently occurs at codon 83, which causes a change of valine to methionine in the MTH1 protein [Wu, C. et al., Biochem. Biophys. Res. Commun. 214 (1995) 1239-1245]. Here we isolated cDNAs for the two types of MTH1 protein and expressed them in Escherichia coli mutT- cells, devoid of their own 8-oxo-dGTPase activity. The two forms of proteins were purified to physical homogeneity, and amino acid analyses confirmed that the variant protein, Met83-MTH1, indeed carries the corresponding amino acid substitution. Met83-MTH1, but not normal type Val83-MTH1, was separated into two peaks in hydrophobic interacting chromatography. 8-Oxo-dGTPase activity of Met83-MTH1 is more thermolabile than that of Val83-MTH1. Circular dichroism (CD) and fluorescence spectroscopic analyses confirmed this conclusion. CD further indicated that Met83-MTH1 has a higher α-helix content. Substitution of valine for methionine at the residue 83 of MYH1 protein appears to lead to alteration in the secondary structure which renders the protein more labile than the normal type protein.

元の言語英語
ページ(範囲)181-194
ページ数14
ジャーナルMutation Research - DNA Repair
384
発行部数3
DOI
出版物ステータス出版済み - 9 1 1997

Fingerprint

Proteins
Substitution reactions
Valine
Circular Dichroism
Methionine
Hydrophobic chromatography
Cells
Amino Acids
Gene encoding
8-oxodGTPase
DNA
Amino Acid Substitution
Nucleosides
Metabolism
Codon
Escherichia coli
Sequence Analysis
Chromatography
Complementary DNA
Fluorescence

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Toxicology
  • Genetics

これを引用

Biochemical and physicochemical characterization of normal and variant forms of human MTH1 protein with antimutagenic activity. / Yakushiji, Hiroyuki; Maraboeuf, Fabrice; Takahashi, Masayuki; Deng, Zeng Sui; Kawabata, Shun-Ichiro; Nakabeppu, Yusaku; Sekiguchi, Mutsuo.

:: Mutation Research - DNA Repair, 巻 384, 番号 3, 01.09.1997, p. 181-194.

研究成果: ジャーナルへの寄稿記事

Yakushiji, Hiroyuki ; Maraboeuf, Fabrice ; Takahashi, Masayuki ; Deng, Zeng Sui ; Kawabata, Shun-Ichiro ; Nakabeppu, Yusaku ; Sekiguchi, Mutsuo. / Biochemical and physicochemical characterization of normal and variant forms of human MTH1 protein with antimutagenic activity. :: Mutation Research - DNA Repair. 1997 ; 巻 384, 番号 3. pp. 181-194.
@article{039a8aefc1834d179701e1042393d152,
title = "Biochemical and physicochemical characterization of normal and variant forms of human MTH1 protein with antimutagenic activity",
abstract = "8-0xo-7,8-dihydro-2'-deoxyguanosine 5'-triphosphate (8-oxo-dGTP) is produced during cellular metabolism, and its misincorporation into DNA causes mutation. Human cells possess an enzyme that hydrolyzes 8-oxo-dGTP to the corresponding nucleoside monophosphate, thereby preventing misincorporation of 8-oxo-7,8-dihydroguanine into DNA. Sequence an analyses of the MTH1 gene, encoding the 8-oxo-7,8-dihydro-2'-deoxyguanosine 5'-triphosphatase (8-oxo-dGTPase) protein in human cell lines revealed that a G to A base substitution frequently occurs at codon 83, which causes a change of valine to methionine in the MTH1 protein [Wu, C. et al., Biochem. Biophys. Res. Commun. 214 (1995) 1239-1245]. Here we isolated cDNAs for the two types of MTH1 protein and expressed them in Escherichia coli mutT- cells, devoid of their own 8-oxo-dGTPase activity. The two forms of proteins were purified to physical homogeneity, and amino acid analyses confirmed that the variant protein, Met83-MTH1, indeed carries the corresponding amino acid substitution. Met83-MTH1, but not normal type Val83-MTH1, was separated into two peaks in hydrophobic interacting chromatography. 8-Oxo-dGTPase activity of Met83-MTH1 is more thermolabile than that of Val83-MTH1. Circular dichroism (CD) and fluorescence spectroscopic analyses confirmed this conclusion. CD further indicated that Met83-MTH1 has a higher α-helix content. Substitution of valine for methionine at the residue 83 of MYH1 protein appears to lead to alteration in the secondary structure which renders the protein more labile than the normal type protein.",
author = "Hiroyuki Yakushiji and Fabrice Maraboeuf and Masayuki Takahashi and Deng, {Zeng Sui} and Shun-Ichiro Kawabata and Yusaku Nakabeppu and Mutsuo Sekiguchi",
year = "1997",
month = "9",
day = "1",
doi = "10.1016/S0921-8777(97)00025-6",
language = "English",
volume = "384",
pages = "181--194",
journal = "Mutation Research - DNA Repair",
issn = "0921-8777",
publisher = "Elsevier BV",
number = "3",

}

TY - JOUR

T1 - Biochemical and physicochemical characterization of normal and variant forms of human MTH1 protein with antimutagenic activity

AU - Yakushiji, Hiroyuki

AU - Maraboeuf, Fabrice

AU - Takahashi, Masayuki

AU - Deng, Zeng Sui

AU - Kawabata, Shun-Ichiro

AU - Nakabeppu, Yusaku

AU - Sekiguchi, Mutsuo

PY - 1997/9/1

Y1 - 1997/9/1

N2 - 8-0xo-7,8-dihydro-2'-deoxyguanosine 5'-triphosphate (8-oxo-dGTP) is produced during cellular metabolism, and its misincorporation into DNA causes mutation. Human cells possess an enzyme that hydrolyzes 8-oxo-dGTP to the corresponding nucleoside monophosphate, thereby preventing misincorporation of 8-oxo-7,8-dihydroguanine into DNA. Sequence an analyses of the MTH1 gene, encoding the 8-oxo-7,8-dihydro-2'-deoxyguanosine 5'-triphosphatase (8-oxo-dGTPase) protein in human cell lines revealed that a G to A base substitution frequently occurs at codon 83, which causes a change of valine to methionine in the MTH1 protein [Wu, C. et al., Biochem. Biophys. Res. Commun. 214 (1995) 1239-1245]. Here we isolated cDNAs for the two types of MTH1 protein and expressed them in Escherichia coli mutT- cells, devoid of their own 8-oxo-dGTPase activity. The two forms of proteins were purified to physical homogeneity, and amino acid analyses confirmed that the variant protein, Met83-MTH1, indeed carries the corresponding amino acid substitution. Met83-MTH1, but not normal type Val83-MTH1, was separated into two peaks in hydrophobic interacting chromatography. 8-Oxo-dGTPase activity of Met83-MTH1 is more thermolabile than that of Val83-MTH1. Circular dichroism (CD) and fluorescence spectroscopic analyses confirmed this conclusion. CD further indicated that Met83-MTH1 has a higher α-helix content. Substitution of valine for methionine at the residue 83 of MYH1 protein appears to lead to alteration in the secondary structure which renders the protein more labile than the normal type protein.

AB - 8-0xo-7,8-dihydro-2'-deoxyguanosine 5'-triphosphate (8-oxo-dGTP) is produced during cellular metabolism, and its misincorporation into DNA causes mutation. Human cells possess an enzyme that hydrolyzes 8-oxo-dGTP to the corresponding nucleoside monophosphate, thereby preventing misincorporation of 8-oxo-7,8-dihydroguanine into DNA. Sequence an analyses of the MTH1 gene, encoding the 8-oxo-7,8-dihydro-2'-deoxyguanosine 5'-triphosphatase (8-oxo-dGTPase) protein in human cell lines revealed that a G to A base substitution frequently occurs at codon 83, which causes a change of valine to methionine in the MTH1 protein [Wu, C. et al., Biochem. Biophys. Res. Commun. 214 (1995) 1239-1245]. Here we isolated cDNAs for the two types of MTH1 protein and expressed them in Escherichia coli mutT- cells, devoid of their own 8-oxo-dGTPase activity. The two forms of proteins were purified to physical homogeneity, and amino acid analyses confirmed that the variant protein, Met83-MTH1, indeed carries the corresponding amino acid substitution. Met83-MTH1, but not normal type Val83-MTH1, was separated into two peaks in hydrophobic interacting chromatography. 8-Oxo-dGTPase activity of Met83-MTH1 is more thermolabile than that of Val83-MTH1. Circular dichroism (CD) and fluorescence spectroscopic analyses confirmed this conclusion. CD further indicated that Met83-MTH1 has a higher α-helix content. Substitution of valine for methionine at the residue 83 of MYH1 protein appears to lead to alteration in the secondary structure which renders the protein more labile than the normal type protein.

UR - http://www.scopus.com/inward/record.url?scp=0030770356&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030770356&partnerID=8YFLogxK

U2 - 10.1016/S0921-8777(97)00025-6

DO - 10.1016/S0921-8777(97)00025-6

M3 - Article

C2 - 9330614

AN - SCOPUS:0030770356

VL - 384

SP - 181

EP - 194

JO - Mutation Research - DNA Repair

JF - Mutation Research - DNA Repair

SN - 0921-8777

IS - 3

ER -