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Kinetics of Methylation by EcoP1I DNA Methyltransferase
Joint Authors
Krishnamurthy, Vinita
Sistla, Srivani
Desirazu, Narasimha Rao
Arathi, Sampath
Bheemanaik, Shivakumara
Source
Issue
Vol. 2010, Issue 2010 (31 Dec. 2010), pp.1-14, 14 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2010-07-15
Country of Publication
Egypt
No. of Pages
14
Main Subjects
Abstract EN
EcoP1I DNA MTase (M.EcoP1I), an N6-adenine MTase from bacteriophage P1, is a part of the EcoP1I restriction-modification (R-M) system which belongs to the Type III R-M system.
It recognizes the sequence 5'-AGACC-3' and methylates the internal adenine.
M.EcoP1I requires Mg2+ for the transfer of methyl groups to DNA.
M.EcoP1I is shown to exist as dimer in solution, and even at high salt concentrations (0.5 M) the dimeric M.EcoP1I does not dissociate into monomers suggesting a strong interaction between the monomer subunits.
Preincubation and isotope partitioning studies with M.EcoP1I indicate a kinetic mechanism where the duplex DNA binds first followed by AdoMet.
Interestingly, M.EcoP1I methylates DNA substrates in the presence of Mn2+ and Ca2+ other than Mg2+ with varying affinities.
Amino acid analysis and methylation assays in the presence of metal ions suggest that M.EcoP1I has indeed two metal ion-binding sites [I358D(x)n…ExK401 and D600xDxD604 motif].
EcoP1I DNA MTase catalyzes the transfer of methyl groups using a distributive mode of methylation on DNA containing more than one recognition site.
A chemical modification of EcoP1I DNA MTase using N-ethylmaleimide resulted in an irreversible inactivation of enzyme activity suggesting the possible role of cysteine residues in catalysis.
American Psychological Association (APA)
Bheemanaik, Shivakumara& Sistla, Srivani& Krishnamurthy, Vinita& Arathi, Sampath& Desirazu, Narasimha Rao. 2010. Kinetics of Methylation by EcoP1I DNA Methyltransferase. Enzyme Research،Vol. 2010, no. 2010, pp.1-14.
https://search.emarefa.net/detail/BIM-461723
Modern Language Association (MLA)
Bheemanaik, Shivakumara…[et al.]. Kinetics of Methylation by EcoP1I DNA Methyltransferase. Enzyme Research No. 2010 (2010), pp.1-14.
https://search.emarefa.net/detail/BIM-461723
American Medical Association (AMA)
Bheemanaik, Shivakumara& Sistla, Srivani& Krishnamurthy, Vinita& Arathi, Sampath& Desirazu, Narasimha Rao. Kinetics of Methylation by EcoP1I DNA Methyltransferase. Enzyme Research. 2010. Vol. 2010, no. 2010, pp.1-14.
https://search.emarefa.net/detail/BIM-461723
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-461723