Site-specific two-color protein labeling for FRET studies using split inteins.
Fluorescence resonance energy transfer (FRET) is an ideal tool for studying protein conformational changes and dynamics. Strategies to specifically label proteins with multiple colors at structurally important sites, a prerequisite for such studies, however, need to be devised on a protein-by-protein basis. Here we describe a simple yet widely applicable two-color site-specific protein labeling method utilizing Npu DnaE split intein.
Yang
Jin-Yi
JY
Institute of Biological Chemistry, Academia Sinica and Institute of Biochemical Sciences, National Taiwan University, Taipei, Taiwan.
Yang
Wei Yuan
WY
D001665:Binding Sites; D003116:Color; D003545:Cysteine; D004258:DNA Polymerase III / Q000737:chemistry / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D047668:Inteins; D046937:Nostoc / Q000201:enzymology; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D019154:Protein Splicing; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*; D013194:Staining and Labeling; D013379:Substrate Specificity
D010446:Peptide Fragments; D011506:Proteins; C074904:DNA polymerase III, alpha subunit; D004258:DNA Polymerase III; D003545:Cysteine