Source:http://linkedlifedata.com/resource/pubmed/id/20814979
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
18
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pubmed:dateCreated |
2010-9-3
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pubmed:abstractText |
Increases in the study of protein-metal complexes, as well as in metal displacement in protein-metal complexes under native conditions for optimum catalytic properties in drug research and catalyst design, demands a separation/detection technology that can accurately measure metal displacement and stoichiometry in protein-metal complexes. Both nuclear magnetic resonance (NMR) and X-ray diffraction techniques have been used for this purpose; however, these techniques lack sensitivity. Electrospray ionization mass spectrometry (ESI-MS) using direct infusion offers higher sensitivity than the former techniques and provides molecular distribution of various protein-metal complexes. However, since protein-metal complexes under native conditions usually are dissolved in salt solutions, their direct ESI-MS analysis requires off-line sample clean-up prior to MS analysis to avoid sample suppression during ESI. Moreover, direct infusion of the salty solution promotes non-specific salt adduct formation by the protein-metal complexes under ESI-MS, which complicates the identification and stoichiometry measurements of the protein-metal complexes. Because of the high mass of protein-metal complexes and lack of sufficient resolution by most mass spectrometers to separate non-specific from specific metal-protein complexes, accurate protein-metal stoichiometry measurements require some form of sample clean up prior to ESI-MS analysis. In this study, we demonstrate that capillary electrophoresis/electrospray ionization in conjunction with a medium-resolution (approximately 10,000) mass spectrometer is an efficient and fast method for the measurement of the stoichiometry of the protein-metal complexes under physiological conditions (pH approximately 7). The metal displacement of Co(2+) to Cd(2+), two metal ions necessary for activation in the monomeric AHL lactonase produced by B. thuringiensis, has been used as a proof of concept.
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pubmed:commentsCorrections | |
pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/AiiA protein, Bacillus,
http://linkedlifedata.com/resource/pubmed/chemical/Bacterial Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Cadmium,
http://linkedlifedata.com/resource/pubmed/chemical/Carboxylic Ester Hydrolases,
http://linkedlifedata.com/resource/pubmed/chemical/Cobalt,
http://linkedlifedata.com/resource/pubmed/chemical/Metalloendopeptidases,
http://linkedlifedata.com/resource/pubmed/chemical/Metalloproteins,
http://linkedlifedata.com/resource/pubmed/chemical/Metals, Heavy,
http://linkedlifedata.com/resource/pubmed/chemical/N-acyl homoserine lactonase
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pubmed:status |
MEDLINE
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pubmed:month |
Sep
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pubmed:issn |
1097-0231
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pubmed:author | |
pubmed:copyrightInfo |
2010 John Wiley & Sons, Ltd.
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pubmed:issnType |
Electronic
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pubmed:volume |
24
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
2730-4
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pubmed:dateRevised |
2011-6-6
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pubmed:meshHeading |
pubmed-meshheading:20814979-Amino Acid Sequence,
pubmed-meshheading:20814979-Bacillus thuringiensis,
pubmed-meshheading:20814979-Bacterial Proteins,
pubmed-meshheading:20814979-Cadmium,
pubmed-meshheading:20814979-Carboxylic Ester Hydrolases,
pubmed-meshheading:20814979-Cobalt,
pubmed-meshheading:20814979-Electrophoresis, Capillary,
pubmed-meshheading:20814979-Metalloendopeptidases,
pubmed-meshheading:20814979-Metalloproteins,
pubmed-meshheading:20814979-Metals, Heavy,
pubmed-meshheading:20814979-Molecular Sequence Data,
pubmed-meshheading:20814979-Spectrometry, Mass, Electrospray Ionization
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pubmed:year |
2010
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pubmed:articleTitle |
Metal displacement and stoichiometry of protein-metal complexes under native conditions using capillary electrophoresis/mass spectrometry.
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pubmed:affiliation |
Cedra Corp., Austin, TX, USA. [corrected]
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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