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(A) Labeling of sfGFP-Y151ThioD with a fluorescein derivative (Fl-cys) by native chemical ligation. The following alkylation reaction was performed in a pH 8 phosphate buffer containing 0.1 mg mL–1 of the above labeling product, 15 mM iodoacetamide, and 10 mM TCEP at RT. (B) SDS-PAGE gel was visualized by Coomassie blue staining (left) and fluorescence emission (right). (C) Mass spectra analysis of the protein variants in this study.
