ArticleProtein science : a publication of the Protein Society2009
A method for site-specific labeling of multiple protein thiols.
Article in Protein science : a publication of the Protein Society, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.
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Who cites it
3 citing papers in PubMed.
- Thiosulfonate-Derived BODIPY "Stick and Glue" Strategy for Fluorescent Thiol Labeling.Angewandte Chemie (International ed. in English) · 2026Article
- Site-directed fluorescence labeling of intrinsically disordered region of human transcription factor YY1: The inhibitory effect of zinc ions.Protein science : a publication of the Protein Society · 2018Article
- Labeling of specific cysteines in proteins using reversible metal protection.Biophysical journal · 2011Article
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Authors and funding
6 authors.
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No grant is acknowledged in the PubMed record.
Abstract
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.
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