Publication Details (including relevant citation information):
Wang, X.; Tipton, J.D.; Emmett, M.R.; Marshall, A.G. Rapid Communications in Mass Spectrometry, 24 (16), 2386-2392, (2010)
Selenomethionine-modified proteins can improve X-ray crystallographic structural resolution by multi-wavelength anomalous diffraction (MAD) phasing. However, the specificity and extent of selenomethionine incorporation must first be assessed. Bottom-up and top-down proteomics with a modified 14.5 T LTQ Fourier transform ion cyclotron resonance mass spectrometer offer a quick, accurate, and robust method to locate and quantify selenomethionine incorporation after auxotrophic expression. Selenomethionine (methionine with sulfur replaced by selenium) has a different natural-abundance isotopic distribution and a mass increase of 47.94 Da relative to wild-type methionine. Here, both wild-type and selenomethionine-substituted forms of the Cas6 protein containing 'clustered regularly interspaced short palindromic repeats' (CRISPRs) were expressed and purified. Comparative bottom-up and top-down proteomics confirmed that all six methionines were fully replaced by selenomethionines in Se-Cas6. Copyright © 2010 John Wiley & Sons, Ltd.
Address (URL): http://onlinelibrary.wiley.com/doi/10.1002/rcm.4655/abstract