Time-resolved fluorescence studies of nucleotide flipping by restriction enzymes
نویسندگان
چکیده
منابع مشابه
Time-resolved fluorescence studies of nucleotide flipping by restriction enzymes
Restriction enzymes Ecl18kI, PspGI and EcoRII-C, specific for interrupted 5-bp target sequences, flip the central base pair of these sequences into their protein pockets to facilitate sequence recognition and adjust the DNA cleavage pattern. We have used time-resolved fluorescence spectroscopy of 2-aminopurine-labelled DNA in complex with each of these enzymes in solution to explore the nucleot...
متن کاملNucleotide flipping by restriction enzymes analyzed by 2-aminopurine steady-state fluorescence
Many DNA modification and repair enzymes require access to DNA bases and therefore flip nucleotides. Restriction endonucleases (REases) hydrolyze the phosphodiester backbone within or in the vicinity of the target recognition site and do not require base extrusion for the sequence readout and catalysis. Therefore, the observation of extrahelical nucleotides in a co-crystal of REase Ecl18kI with...
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DNA base flipping is an important mechanism in molecular enzymology, but its study is limited by the lack of an accessible and reliable diagnostic technique. A series of crystalline complexes of a DNA methyltransferase, M.HhaI, and its cognate DNA, in which a fluorescent nucleobase analogue, 2-aminopurine (AP), occupies defined positions with respect the target flipped base, have been prepared ...
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ژورنال
عنوان ژورنال: Nucleic Acids Research
سال: 2009
ISSN: 1362-4962,0305-1048
DOI: 10.1093/nar/gkp688