title: Single-Molecule Imaging With One Color Fluorescence.
abstract: Single-molecule fluorescence imaging is a powerful tool that enables real-time observation of DNA-protein or RNA-protein interactions with a nanometer precision. Here, we provide a detailed procedure for a previously developed single-molecule fluorescence method, termed "single-molecule protein-induced fluorescence enhancement" (smPIFE). While smFRET (Förster resonance energy transfer) requires both donor and acceptor, protein-induced fluorescence enhancement (PIFE) employs a single dye and measures the increase in fluorescence intensity induced by protein binding near the dye. PIFE displays distance sensitivity within 0-4nm, making it a powerful complementary or alternative tool to FRET method. In this chapter, we will discuss the various ways that PIFE has been utilized to study protein-nucleic acid interactions.
authors: Johns Hopkins University, Baltimore, MD, United States.
authors: Y
authors: Y
authors: Qiu
authors: Johns Hopkins University, Baltimore, MD, United States. Electronic address: smyong@jhu.edu.
authors: S
authors: S
authors: Myong
mesh_terms: D004247:DNA / Q000737:chemistry / Q000302:isolation & purification*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification*; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry; D057054:Molecular Imaging / Q000379:methods*; D011485:Protein Binding; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification*; D013050:Spectrometry, Fluorescence / Q000379:methods
chemical_list: D004268:DNA-Binding Proteins; D005456:Fluorescent Dyes; D016601:RNA-Binding Proteins; D004247:DNA
keywords: Förster resonance energy transfer; One-color imaging; Protein–nucleic acid interactions; Single-molecule protein-induced fluorescence enhancement