Linked Life Data

19153692

Source:http://linkedlifedata.com/resource/pubmed/id/19153692

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:dateCreated
2009-1-20
pubmed:abstractText
The modification of cell surface proteins by plasma membrane and soluble proteases is important for physiological and pathological processes. Methods to identify shed and soluble substrates are crucial to further define the substrate repertoire, termed the substrate degradome, of individual proteases. Identifying protease substrates is essential to elucidate protease function and involvement in different homeostatic and disease pathways. This characterisation is also crucial for drug target identification and validation, which would then allow the rational design of specific targeted inhibitors for therapeutic intervention. We describe two methods for identifying and quantifying shed cell surface protease targets in cultured cells utilising Isotope-Coded Affinity Tags (ICAT) and Isobaric Tags for Relative and Absolute Quantification (iTRAQ). As a model system to develop these techniques, we chose a cell-membrane expressed matrix metalloproteinase, MMP-14, but the concepts can be applied to proteases of other classes. By over-expression, or conversely inhibition, of a particular protease with careful selection of control conditions (e.g. vector or inactive protease) and differential labelling, shed proteins can be identified and quantified by mass spectrometry (MS), MS/MS fragmentation and database searching.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:issn
1064-3745
pubmed:author
pubmed:issnType
Print
pubmed:volume
528
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
159-76
pubmed:meshHeading
pubmed:year
2009
pubmed:articleTitle
Membrane protease degradomics: proteomic identification and quantification of cell surface protease substrates.
pubmed:affiliation
Centre for Blood Research, University of British Columbia, Vancouver, BC, Canada.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't, Validation Studies