O6-Methylguanine-DNA methyltransferase of human lymphoid cells: structural and kinetic properties and absence in repair-deficient cells.
Harris AL., Karran P., Lindahl T.
Human lymphoid cell lines contain a DNA repair enzyme which removes the mutagenic alkylation lesion O6-methylguanine from DNA. The enzyme transfers the methyl group to a protein cysteine residue, generating S-methylcysteine, and is inactivated as a consequence of the reaction. Apparently the methylated enzyme represents a dead-end complex. The transfer reaction is very rapid and is completed in less than 1 min at 37 degrees, but methyl group transfer from single-stranded DNA or heavily damaged DNA is less efficient. The active methyltransferase and the methylated protein both have molecular weights of 21,000 to 22,000, as determined by gel filtration. Lymphoid cell lines proficient in repair of O6-methylguanine in vivo, Mex+, contain 10,000 to 25,000 molecules of the methyltransferase per cell. In contrast, repair-deficient cell lines, Mex-, do not contain detectable amounts of the enzyme. The latter point was verified by applying a partial purification procedure for the enzyme to cell-free extracts from two Mex- cell lines.