Source:http://linkedlifedata.com/resource/pubmed/id/15262447
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
2
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| pubmed:dateCreated |
2004-7-20
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| pubmed:abstractText |
Electron spin resonance (ESR) spectroscopy in combination with site-directed spin labeling (SDSL) is a powerful tool for determining protein structure, dynamics and interactions. We report here a method for determining interactions between spin labels and paramagnetic relaxation agents, which is performed under subsaturating conditions. The low microwave-field amplitude employed (h(1)<0.36 G) only requires standard, commercially available ESR equipment. The effect of relaxation enhancement on the spin-spin-relaxation time, T(2e), is measured by this method, and compared to classical progressive power saturation performed on a free spin label, (1-oxyl-2,2,5,5-tetramethyl-Delta(3)-pyrroline-3-methyl)methanethiosulfonate (MTSL), and a spin-labeled protein (Thermomyces lanuginosa lipase, TLL-I252C), employing the water-soluble relaxation agent chromium(III) oxalate (Crox) in concentrations between 0-10 mM. The low-amplitude theory showed excellent agreement with that of classical power saturation in quantifying Crox-induced relaxation enhancement. Low-amplitude measurements were then performed using a standard resonator, with Crox, on 11 spin-labeled TLL mutants displaying rotational correlation times in the motional narrowing regime. All spin-labeled proteins exhibited significant changes in T(2e). We postulate that this novel method is especially suitable for studying moderately immobilized spin labels, such as those positioned at exposed sites in a protein. This method should prove useful for research groups with access to any ESR instrumentation.
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| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/(1-oxyl-2,2,5,5-tetramethylpyrroline...,
http://linkedlifedata.com/resource/pubmed/chemical/Cyclic N-Oxides,
http://linkedlifedata.com/resource/pubmed/chemical/Cysteine,
http://linkedlifedata.com/resource/pubmed/chemical/Lipase,
http://linkedlifedata.com/resource/pubmed/chemical/Mesylates,
http://linkedlifedata.com/resource/pubmed/chemical/Peptides,
http://linkedlifedata.com/resource/pubmed/chemical/Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Spin Labels,
http://linkedlifedata.com/resource/pubmed/chemical/Water
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| pubmed:status |
MEDLINE
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| pubmed:month |
Aug
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| pubmed:issn |
0165-022X
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| pubmed:author | |
| pubmed:issnType |
Print
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| pubmed:day |
31
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| pubmed:volume |
60
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
117-38
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| pubmed:dateRevised |
2006-11-15
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| pubmed:meshHeading |
pubmed-meshheading:15262447-Ascomycota,
pubmed-meshheading:15262447-Biophysics,
pubmed-meshheading:15262447-Cyclic N-Oxides,
pubmed-meshheading:15262447-Cysteine,
pubmed-meshheading:15262447-Dose-Response Relationship, Drug,
pubmed-meshheading:15262447-Electron Spin Resonance Spectroscopy,
pubmed-meshheading:15262447-Lipase,
pubmed-meshheading:15262447-Magnetic Resonance Spectroscopy,
pubmed-meshheading:15262447-Mesylates,
pubmed-meshheading:15262447-Microwaves,
pubmed-meshheading:15262447-Mutagenesis, Site-Directed,
pubmed-meshheading:15262447-Mutation,
pubmed-meshheading:15262447-Peptides,
pubmed-meshheading:15262447-Proteins,
pubmed-meshheading:15262447-Recombinant Proteins,
pubmed-meshheading:15262447-Spin Labels,
pubmed-meshheading:15262447-Time Factors,
pubmed-meshheading:15262447-Water
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| pubmed:year |
2004
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| pubmed:articleTitle |
Low microwave-amplitude ESR spectroscopy: measuring spin-relaxation interactions of moderately immobilized spin labels in proteins.
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| pubmed:affiliation |
Department of Biotechnology, Royal Institute of Technology, AlbaNova University Center, SE-106 91, Stockholm, Sweden. eva@biochem.kth.se
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| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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