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Biophys Chem ; 103(2): 129-37, 2003 Jan 21.
Article in English | MEDLINE | ID: mdl-12568936

ABSTRACT

The methyltransferase component of type I DNA restriction and modification systems comprises three subunits, one DNA sequence specificity subunit and two DNA modification subunits. Limited proteolysis of the EcoKI methyltransferase shows that a 55-kDa N-terminal fragment of the 59-kDa modification subunit is resistant to degradation. We have purified this fragment and determined by mass spectrometry that proteolysis removes 43 or 44 amino acids from the C-terminus. The fragment fails to interact with the other subunits even though it still possesses secondary and tertiary structure and the ability to bind the S-adenosylmethionine cofactor. We conclude that the C-terminal region of the modification subunit of EcoKI is essential for the assembly of the EcoKI methyltransferase.


Subject(s)
Protein Subunits/chemistry , Site-Specific DNA-Methyltransferase (Adenine-Specific)/biosynthesis , Amino Acid Sequence , Peptide Fragments/chemistry , Peptide Fragments/isolation & purification , Peptide Fragments/metabolism , Peptide Mapping , Protein Denaturation , Protein Folding , Protein Structure, Secondary , Protein Structure, Tertiary , Protein Subunits/metabolism , Site-Specific DNA-Methyltransferase (Adenine-Specific)/chemistry
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