Linked Life Data

18756552

Source:http://linkedlifedata.com/resource/pubmed/id/18756552

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
14
pubmed:dateCreated
2008-9-26
pubmed:abstractText
The selective modification of proteins with a synthetic probe is of central interest for many aspects of protein chemistry. We have recently reported a new approach in which a short cysteine-containing tag (CysTag) fused to one part of a split intein is first modified with a sulfhydryl-reactive probe. In a second step, protein trans-splicing is used to link the labelled CysTag to a target protein that has been expressed in fusion with the complementary split intein fragment. Here, we present the generation and biochemical characterisation of the artificially split Mycobacterium xenopi GyrA intein. We show that this split intein is active without a renaturation step and that it provides a significant improvement for the CysTag protein-labelling approach in terms of product yields and target protein tolerance. Two proteins with multiple cysteine residues, human growth hormone and a multidomain nonribosomal peptide synthetase, were site-specifically modified with high yields. Our approach combines the benefits of the plethora of commercially available cysteine-reactive probes with a straightforward route for their site-specific incorporation even into complex and cysteine-rich proteins.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Sep
pubmed:issn
1439-7633
pubmed:author
pubmed:issnType
Electronic
pubmed:day
22
pubmed:volume
9
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
2317-25
pubmed:meshHeading
pubmed:year
2008
pubmed:articleTitle
Site-specific chemical modification of proteins with a prelabelled cysteine tag using the artificially split Mxe GyrA intein.
pubmed:affiliation
Technische Universität Dortmund, Fakultät Chemie-Chemische Biologie, Otto-Hahn-Strasse 6, 44229 Dortmund, Germany.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't