Source:http://linkedlifedata.com/resource/pubmed/id/15905179
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
8
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pubmed:dateCreated |
2005-8-15
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pubmed:abstractText |
Here we describe an original strategy for unbiased quantification of protein expression called difference in mass analysis using labeled lysine (K) (DIMAL-K). DIMAL-K is based on the differential predigestion labeling of lysine residues in complex protein mixtures. The method is relevant for proteomic analysis by two-dimensional electrophoresis and MALDI-TOF mass spectrometry. Protein labeling on lysine residues uses two closely related chemical reagents, S-methyl thioacetimidate and S-methyl thiopropionimidate. Using protein standards, we demonstrated that 1) the chemical labeling was quantitative, specific, and rapid; 2) the differentially labeled proteins co-migrated on two-dimensional gels; and 3) the identification by mass fingerprinting and the relative quantification of the proteins were possible from a single MALDI-TOF mass spectrum. The power of the method was tested by comparing and quantifying the secretion of proteins in normal and proinflammatory astrocytic secretomes (20 microg). We showed that DIMAL-K was more sensitive and accurate than densitometric image analysis and allowed the detection and quantification of novel proteins.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Aug
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pubmed:issn |
1535-9476
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
4
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
1085-94
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:15905179-Animals,
pubmed-meshheading:15905179-Astrocytes,
pubmed-meshheading:15905179-Cells, Cultured,
pubmed-meshheading:15905179-Electrophoresis, Gel, Two-Dimensional,
pubmed-meshheading:15905179-Isotope Labeling,
pubmed-meshheading:15905179-Lysine,
pubmed-meshheading:15905179-Mice,
pubmed-meshheading:15905179-Peptide Fragments,
pubmed-meshheading:15905179-Proteome,
pubmed-meshheading:15905179-Spectrometry, Mass, Matrix-Assisted Laser...
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pubmed:year |
2005
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pubmed:articleTitle |
Difference in mass analysis using labeled lysines (DIMAL-K): a new, efficient proteomic quantification method applied to the analysis of astrocytic secretomes.
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pubmed:affiliation |
Département de Neurobiologie, Institut de Génomique Fonctionnelle, 141 rue de la cardonille, 34094 Montpellier Cedex 5, France.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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