2c7r

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{{STRUCTURE_2c7r| PDB=2c7r | SCENE= }}
{{STRUCTURE_2c7r| PDB=2c7r | SCENE= }}
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'''HHAI DNA METHYLTRANSFERASE (T250G MUTANT) COMPLEX WITH OLIGONUCLEOTIDE CONTAINING 2-AMINOPURINE AS A TARGET BASE (GPGC:GMGC) AND SAH'''
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===HHAI DNA METHYLTRANSFERASE (T250G MUTANT) COMPLEX WITH OLIGONUCLEOTIDE CONTAINING 2-AMINOPURINE AS A TARGET BASE (GPGC:GMGC) AND SAH===
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==Overview==
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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 and their structures determined at higher than 2 A resolution. From time-resolved fluorescence measurements of these single crystals, we have established that the fluorescence decay function of AP shows a pronounced, characteristic response to base flipping: the loss of the very short (approximately 100 ps) decay component and the large increase in the amplitude of the long (approximately 10 ns) component. When AP is positioned at sites other than the target site, this response is not seen. Most significantly, we have shown that the same clear response is apparent when M.HhaI complexes with DNA in solution, giving an unambiguous signal of base flipping. Analysis of the AP fluorescence decay function reveals conformational heterogeneity in the DNA-enzyme complexes that cannot be discerned from the present X-ray structures.
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{{ABSTRACT_PUBMED_16340006}}
==About this Structure==
==About this Structure==
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[[Category: Restriction system]]
[[Category: Restriction system]]
[[Category: Transferase]]
[[Category: Transferase]]
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Revision as of 06:56, 29 July 2008

Image:2c7r.png

Template:STRUCTURE 2c7r

HHAI DNA METHYLTRANSFERASE (T250G MUTANT) COMPLEX WITH OLIGONUCLEOTIDE CONTAINING 2-AMINOPURINE AS A TARGET BASE (GPGC:GMGC) AND SAH

Template:ABSTRACT PUBMED 16340006

About this Structure

2C7R is a Protein complex structure of sequences from Haemophilus haemolyticus. Full crystallographic information is available from OCA.

Reference

Time-resolved fluorescence of 2-aminopurine as a probe of base flipping in M.HhaI-DNA complexes., Neely RK, Daujotyte D, Grazulis S, Magennis SW, Dryden DT, Klimasauskas S, Jones AC, Nucleic Acids Res. 2005 Dec 9;33(22):6953-60. Print 2005. PMID:16340006

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