Linked Life Data

21604133

Source:http://linkedlifedata.com/resource/pubmed/id/21604133

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:dateCreated
2011-5-23
pubmed:abstractText
This paper describes a cyclic on-column procedure for the sequential degradation of complex O-glycans on proteins by periodate oxidation of sugars and cleavage of oxidation products by elimination. Glycoproteins are immobilized to alkali-stable, reversed-phase Poros 20 beads, desialylated by treatment with dilute trifluoroacetic acid, and de-O-glycosylated by two degradation cycles before the eluted apoproteins are digested with trypsin for analysis by liquid chromatography electrospray ionization-mass spectrometry. Even complex glycan moieties are removed under mild conditions with only minimal effects on structural integrity of the peptide core by fragmentation, dehydration, or racemization of lysine and arginine residues. The protocol is also applicable on gel-immobilized glycoproteins after 1D or 2D gel electrophoresis. Conversion of O-glycoproteins into their corresponding apoproteins results in facilitated accessibility of tryptic cleavage sites, increases the numbers of peptide fragments, and accordingly enhances protein coverage and identification rates within the subproteome of mucin-type O-glycoproteins. The protocol is suitable for automatization, but due to partial elution from the Poros 20 columns it is not recommended for applications on the glycopeptide level.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:issn
1940-6029
pubmed:author
pubmed:issnType
Electronic
pubmed:volume
753
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
323-33
pubmed:meshHeading
pubmed:year
2011
pubmed:articleTitle
Chemical de-O-glycosylation of glycoproteins for applications in LC-based proteomics.
pubmed:affiliation
Institute of Biochemistry II, Medical Faculty, Center for Molecular Medicine Cologne, University of Cologne, 50931, Köln, Germany. franz.hanisch@uni-koeln.de
pubmed:publicationType
Journal Article