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Fig. 3 Selective labeling of a Sec-containing EGFP using a maleimide
probe. HPLC/ESI-MS analysis of an equimolar mixture of DMNB-caged
EGFPTyr39Sec and EGFPTyr39Cys proteins (upper) and following its
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previously described for any protein or a synthetic peptide.31
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and practical utility of the new technique. The generality of this
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HpaII DNA cytosine-5 methyltransferase32,33 fusion protein
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We are grateful to Peter G. Schultz and Lei Wang for their
kind gifts of the LWUPF1D strain and associated plasmids. This
work was supported by the Research Council of Lithuania
(grant MIP-115/2012) and the European Social Fund under
National Integrated Programme Biotechnology & Biopharmacy
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Notes and references
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§ Minor peaks to the right from the major peaks manifest both 35 R. K. Neely, P. Dedecker, J. I. Hotta, G. Urbanaviciu¯te, S. Klimasauskas
inherent and apparent heterogeneity of the EGFP proteins in solution,
and J. Hofkens, Chem. Sci., 2010, 1, 453–460.
which comprises inadvertent oxidation (increments of +16 amu) and/or 36 Z. Liutkeviˇciu¯t˙e, E. Kriukien˙e, I. Grigaityt˙e, V. Maseviˇcius and
bound counterions (Na+, K+). Oxidation of side chains (Met, His, Cys) in
S. Klimasauskas, Angew. Chem., Int. Ed. Engl., 2011, 50, 2090–2093.
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proteins commonly occurs to different degrees during biosynthesis and 37 Z. Liutkeviciu¯te, E. Kriukiene, J. Licyte, M. Rudyte, G. Urbanaviciu¯te
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handling or due to ESI-MS artifacts under aerobic conditions.
and S. Klimasauskas, J. Am. Chem. Soc., 2014, 136, 5884–5887.
1 E. M. Brustad and F. H. Arnold, Curr. Opin. Chem. Biol., 2011, 15,
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