Purification and biochemical characterization of the EcaI DNA methyltransferase

L. Szilák, P. Venetianer, A. Kiss

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

The EcaI GGTNACC-specific DNA-adenine modification methyltransferase has been purified to apparent homogeneity. The active form of the DNA methyltransferase is a single polypeptide. The enzyme has a pH optimum at pH 8.0 and a temperature optimum at 25°C. EcaI DNA methyltransferase transfers one methyl group to the adenine of the recognition site in a single binding event. The K(m) was 170 nM for DNA and 1.8 μM for the methyl donor S-adenosylmethionine. Methylated DNA is a competitive inhibitor with respect to DNA (K(i) = 3.5 nM). The other product of the DNA-methylation reaction, S-adenosylhomocysteine was found to be a competitive inhibitor with respect to S-adenosylmethionine (K(i) = 2.7 μM). The S-adenosylmethionine analog sinefungin was shown to be a very strong inhibitor (K(i) = 3.5 nM) of the DNA methyltransferase reaction.

Original languageEnglish
Pages (from-to)391-397
Number of pages7
JournalEuropean Journal of Biochemistry
Volume209
Issue number1
DOIs
Publication statusPublished - 1992

Fingerprint

Purification
S-Adenosylmethionine
DNA
Methyltransferases
sinefungin
Adenine
DNA Modification Methylases
S-Adenosylhomocysteine
DNA Methylation
DNA modification methylase EcaI
Peptides
Temperature
Enzymes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Purification and biochemical characterization of the EcaI DNA methyltransferase. / Szilák, L.; Venetianer, P.; Kiss, A.

In: European Journal of Biochemistry, Vol. 209, No. 1, 1992, p. 391-397.

Research output: Contribution to journalArticle

@article{734bbc63e5ae475da9b21b28bebe40ac,
title = "Purification and biochemical characterization of the EcaI DNA methyltransferase",
abstract = "The EcaI GGTNACC-specific DNA-adenine modification methyltransferase has been purified to apparent homogeneity. The active form of the DNA methyltransferase is a single polypeptide. The enzyme has a pH optimum at pH 8.0 and a temperature optimum at 25°C. EcaI DNA methyltransferase transfers one methyl group to the adenine of the recognition site in a single binding event. The K(m) was 170 nM for DNA and 1.8 μM for the methyl donor S-adenosylmethionine. Methylated DNA is a competitive inhibitor with respect to DNA (K(i) = 3.5 nM). The other product of the DNA-methylation reaction, S-adenosylhomocysteine was found to be a competitive inhibitor with respect to S-adenosylmethionine (K(i) = 2.7 μM). The S-adenosylmethionine analog sinefungin was shown to be a very strong inhibitor (K(i) = 3.5 nM) of the DNA methyltransferase reaction.",
author = "L. Szil{\'a}k and P. Venetianer and A. Kiss",
year = "1992",
doi = "10.1111/j.1432-1033.1992.tb17301.x",
language = "English",
volume = "209",
pages = "391--397",
journal = "FEBS Journal",
issn = "1742-464X",
publisher = "Wiley-Blackwell",
number = "1",

}

TY - JOUR

T1 - Purification and biochemical characterization of the EcaI DNA methyltransferase

AU - Szilák, L.

AU - Venetianer, P.

AU - Kiss, A.

PY - 1992

Y1 - 1992

N2 - The EcaI GGTNACC-specific DNA-adenine modification methyltransferase has been purified to apparent homogeneity. The active form of the DNA methyltransferase is a single polypeptide. The enzyme has a pH optimum at pH 8.0 and a temperature optimum at 25°C. EcaI DNA methyltransferase transfers one methyl group to the adenine of the recognition site in a single binding event. The K(m) was 170 nM for DNA and 1.8 μM for the methyl donor S-adenosylmethionine. Methylated DNA is a competitive inhibitor with respect to DNA (K(i) = 3.5 nM). The other product of the DNA-methylation reaction, S-adenosylhomocysteine was found to be a competitive inhibitor with respect to S-adenosylmethionine (K(i) = 2.7 μM). The S-adenosylmethionine analog sinefungin was shown to be a very strong inhibitor (K(i) = 3.5 nM) of the DNA methyltransferase reaction.

AB - The EcaI GGTNACC-specific DNA-adenine modification methyltransferase has been purified to apparent homogeneity. The active form of the DNA methyltransferase is a single polypeptide. The enzyme has a pH optimum at pH 8.0 and a temperature optimum at 25°C. EcaI DNA methyltransferase transfers one methyl group to the adenine of the recognition site in a single binding event. The K(m) was 170 nM for DNA and 1.8 μM for the methyl donor S-adenosylmethionine. Methylated DNA is a competitive inhibitor with respect to DNA (K(i) = 3.5 nM). The other product of the DNA-methylation reaction, S-adenosylhomocysteine was found to be a competitive inhibitor with respect to S-adenosylmethionine (K(i) = 2.7 μM). The S-adenosylmethionine analog sinefungin was shown to be a very strong inhibitor (K(i) = 3.5 nM) of the DNA methyltransferase reaction.

UR - http://www.scopus.com/inward/record.url?scp=0026706564&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026706564&partnerID=8YFLogxK

U2 - 10.1111/j.1432-1033.1992.tb17301.x

DO - 10.1111/j.1432-1033.1992.tb17301.x

M3 - Article

VL - 209

SP - 391

EP - 397

JO - FEBS Journal

JF - FEBS Journal

SN - 1742-464X

IS - 1

ER -