Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
9
pubmed:dateCreated
2010-9-3
pubmed:abstractText
Although enzyme catalyzed 18O labeling has been used as a tool in quantitative proteomics, this type of labeling has not yielded the same impact yet as alternative techniques for quantitation like SILAC or labeling with chemical mass tags. The practical difficulties involved in 18O labeling, most importantly the occurrence of incomplete labeling and, as a result, the difficulties in data analysis and interpretation have hampered its implementation in high-throughput comparative proteomics protocols. In this paper, we have optimized the 18O labeling procedure to such an extent that complete labeling can be achieved in a routine manner. We have implemented this approach into a protein-protein interaction analysis pipeline to differentiate between bona fide interaction partners of the low-level expressing cell cycle regulator cyclin-dependent kinase 9 (Cdk9) and nonspecifically binding or background proteins. Previously known as well as novel interaction partners of Cdk9 were found, among which most notably the Mediator complex and several other proteins involved in transcriptional regulation. We show here that a differential proteomics approach based on 18O labeling provides a valuable method for high-confidence determination of protein interaction partners and is easily implemented in protein network analysis workflows.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Sep
pubmed:issn
1535-3907
pubmed:author
pubmed:issnType
Electronic
pubmed:day
3
pubmed:volume
9
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
4464-75
pubmed:meshHeading
pubmed:year
2010
pubmed:articleTitle
Differential proteomics based on 18O labeling to determine the cyclin dependent kinase 9 interactome.
pubmed:affiliation
Proteomics Center and Department of Cell Biology, Erasmus University Medical Center, Dr. Molewaterplein 50, 3015 GE Rotterdam, The Netherlands.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't