Linked Life Data

19388048

Source:http://linkedlifedata.com/resource/pubmed/id/19388048

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
5
pubmed:dateCreated
2009-4-29
pubmed:abstractText
We present a generic method for the site-specific and differential labeling of multiple cysteine residues in one protein. Phenyl arsenic oxide has been employed as a protecting group of two closely spaced thiols, allowing first labeling of a single thiol. Subsequently, the protecting group is removed, making available a reactive dithiol site for labeling with a second probe. For proof-of-principle, single and triple Cys mutants of the sulphate binding protein of an ABC transporter were constructed. The closely spaced thiols were engineered on the basis of the crystal structure of the protein and placed in different types of secondary structure elements and at different spacing. We show that phenyl arsenic oxide is a good protecting group for thiols spaced 6.3-7.3 A. Proteins were labeled with two different fluorescent labels and the labeling ratios were determined with UV-Vis spectroscopy and MALDI-Tof mass spectrometry. The average labeling efficiency was approximately 80% for the single thiol and 65-90% for the dithiol site.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/19388048-10545166, http://linkedlifedata.com/resource/pubmed/commentcorrection/19388048-11101244, http://linkedlifedata.com/resource/pubmed/commentcorrection/19388048-11533486, http://linkedlifedata.com/resource/pubmed/commentcorrection/19388048-14751262, http://linkedlifedata.com/resource/pubmed/commentcorrection/19388048-15034592, http://linkedlifedata.com/resource/pubmed/commentcorrection/19388048-16375896, http://linkedlifedata.com/resource/pubmed/commentcorrection/19388048-16748050, http://linkedlifedata.com/resource/pubmed/commentcorrection/19388048-17313455, http://linkedlifedata.com/resource/pubmed/commentcorrection/19388048-3288756, http://linkedlifedata.com/resource/pubmed/commentcorrection/19388048-5333560, http://linkedlifedata.com/resource/pubmed/commentcorrection/19388048-5638587, http://linkedlifedata.com/resource/pubmed/commentcorrection/19388048-8563639
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
May
pubmed:issn
1469-896X
pubmed:author
pubmed:issnType
Electronic
pubmed:volume
18
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1033-41
pubmed:dateRevised
2010-9-27
pubmed:meshHeading
pubmed:year
2009
pubmed:articleTitle
A method for site-specific labeling of multiple protein thiols.
pubmed:affiliation
Department of Biochemistry, Groningen Biomolecular Science and Biotechnology Institute & Zernike Institute for Advanced Materials, University of Groningen, 9747 AG Groningen, The Netherlands.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't