The kinetic mechanism of phage T4 DNA-[N6-adenine]-methyltransferase

被引：3

作者：

Evdokimov, AA ^{[1
]}

Zinoviev, VV ^{[1
]}

Malygin, EG ^{[1
]}

机构：

[1] State Res Ctr Virol & Biotechnol VECTOR, Inst Mol Biol, Koltsov 633159, Novosibirsk Reg, Russia

来源：

MOLECULAR BIOLOGY | 2002年 / 36卷 / 05期

基金：

俄罗斯基础研究基金会;

关键词：

DNA methyltransferase; oligodeoxyribonucleotides; steady-state kinetics; enzyme isomerization; sequential mechanism;

D O I：

10.1023/A:1020627514839

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Kinetic analysis of methyl group transfer from S-adenosyl-L-methionine (SAM) to the GATC recognition site catalyzed by the phage T4 DNA-[N6-adenine]-methyltransferase (MTase) [EC 2.1.1.72] showed that the reverse reaction is at least 500 times slower than the direct one. The overall pattern product inhibition corresponds to an ordered steady-state mechanism following the sequence SAMdown arrowDNAdown arrowmetDNAup arrowSAHup arrow (S-adenosyl-L-homocysteine). Pronounced inhibition was observed at high concentrations of the 20-meric substrate duplex, which may be attributed to formation of a dead-end complex MTase-SAH-DNA. In contrast, high SAM concentrations proportionally accelerated the reaction. Thus, the reaction may include a stage whereby the binding of SAM and the release of SAH are united into one concerted event. Computer fitting of alternative kinetic schemes to the aggregate of experimental data revealed that the most plausible mechanism involves isomerization of the enzyme.

引用

页码：683 / 692

页数：10

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