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Insights into copper coordination in the EcoRI–DNA complex by ESR spectroscopy

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Insights into copper coordination in the EcoRI–DNA complex by ESR spectroscopy

Auteurs : Ming Ji [États-Unis] ; Likun Tan [États-Unis] ; Linda Jen-Jacobson [États-Unis] ; Sunil Saxena [États-Unis]

Source :

Molecular physics [ 0026-8976 ] ; 2014.

RBID : PMC:4350447

Abstract

The EcoRI restriction endonuclease requires one divalent metal ion in each of two symmetrical and identical catalytic sites to catalyse double-strand DNA cleavage. Recently, we showed that Cu²⁺ binds outside the catalytic sites to a pair of new sites at H114 in each sub-unit, and inhibits Mg²⁺ -catalysed DNA cleavage. In order to provide more detailed structural information on this new metal ion binding site, we performed W-band (~94 GHz) and X-band (~9.5 GHz) electron spin resonance spectroscopic measurements on the EcoRI–DNA–(Cu²⁺ )₂ complex. Cu²⁺ binding results in two distinct components with different g_zz and A_zz values. X-band electron spin echo envelope modulation results indicate that both components arise from a Cu²⁺ coordinated to histidine. This observation is further confirmed by the hyperfine sub-level correlation results. W-band electron nuclear double resonance spectra provide evidence for equatorial coordination of water molecules to the Cu²⁺ ions.

Url:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4350447

DOI: 10.1080/00268976.2014.934313
PubMed: 25750461
PubMed Central: 4350447

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PMC:4350447

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<front><div type="abstract" xml:lang="en"><p id="P1">The EcoRI restriction endonuclease requires one divalent metal ion in each of two symmetrical and identical catalytic sites to catalyse double-strand DNA cleavage. Recently, we showed that Cu<sup>2+</sup>
 binds outside the catalytic sites to a pair of new sites at H114 in each sub-unit, and inhibits Mg<sup>2+</sup>
 -catalysed DNA cleavage. In order to provide more detailed structural information on this new metal ion binding site, we performed W-band (~94 GHz) and X-band (~9.5 GHz) electron spin resonance spectroscopic measurements on the EcoRI–DNA–(Cu<sup>2+</sup>
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 complex. Cu<sup>2+</sup>
 binding results in two distinct components with different <italic>g</italic>
<sub>zz</sub>
 and <italic>A</italic>
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 values. X-band electron spin echo envelope modulation results indicate that both components arise from a Cu<sup>2+</sup>
 coordinated to histidine. This observation is further confirmed by the hyperfine sub-level correlation results. W-band electron nuclear double resonance spectra provide evidence for equatorial coordination of water molecules to the Cu<sup>2+</sup>
 ions.</p>
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Insights into copper coordination in the EcoRI–DNA complex by ESR spectroscopy

Insights into copper coordination in the EcoRI–DNA complex by ESR spectroscopy

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Abstract

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