Energy Migration and Fluorescence Depolarization: Structural Studies of Ethidium Bromide-Nucleic Acid Complexes

1983 ◽  
pp. 523-539
Author(s):  
D. Genest ◽  
Ph. Wahl
Keyword(s):  
Nucleic Acid ◽  
Ethidium Bromide ◽  
Energy Migration ◽  
Structural Studies ◽  
Fluorescence Depolarization

Comparative analysis of the DNA staining efficiencies of different fluorescent dyes in preparative agarose gel electrophoresis

2005 ◽  
Vol 43 (8) ◽  
Author(s):  
Qing Huang ◽  
Wei-Ling Fu
Keyword(s):  
Comparative Analysis ◽  
Nucleic Acid ◽  
Nucleic Acids ◽  
Ethidium Bromide ◽  
Gel Electrophoresis ◽  
Fluorescent Dyes ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Dna Staining

AbstractEthidium bromide (EB) is a mutagen and toxin that is widely used in the laboratory for visualization of nucleic acids. Safer nucleic acid stains, such as SYBR

scholarly journals Label-free horizontal EMSA for analysis of protein-RNA interactions

2019 ◽  
Author(s):  
William Perea ◽  
Nancy L. Greenbaum
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Dissociation Constant ◽  
Intermolecular Interactions ◽  
Ethidium Bromide ◽  
Fluorescent Labeling ◽  
Label Free ◽  
Cationic Protein ◽  
Intercalating Dye ◽  
Protein Components

AbstractWe describe a method to analyze the affinity and specificity of interactions between proteins and RNA using horizontal PAGE under non-denaturing conditions. The method permits tracking of migration of anionic and cationic biomolecules and complexes toward anode and cathode, respectively, therefore enabling quantification of bound and free biomolecules of different charges and affinity of their intermolecular interactions. The gel is stained with a fluorescent intercalating dye (SYBR®Gold or ethidium bromide) for visualization of nucleic acids followed by Coomassie® Brilliant Blue R-250 for visualizations of proteins; the dissociation constant is determined separately from the intensity of unshifted and shifted bands visualized by each dye. The method permits calculation of bound and unbound anionic nucleic acid and cationic protein components in the same gel, regardless of charge, under identical conditions, and avoids the need for radioisotope or fluorescent labeling of either component.

scholarly journals Molecular mechanisms of substrate-controlled ring dynamics and sub-stepping in a nucleic-acid dependent hexameric motor

2016 ◽  
Author(s):  
Nathan D. Thomsen ◽  
Michael R. Lawson ◽  
Lea B. Witkowsky ◽  
Song Qu ◽  
James M. Berger
Keyword(s):  
Nucleic Acid ◽  
Conformational Changes ◽  
Molecular Motors ◽  
Structural Changes ◽  
Molecular Mechanisms ◽  
Atp Hydrolysis ◽  
Ring Closure ◽  
Structural Studies ◽  
Saxs Data ◽  
Structural Methods

ABSTRACTRing-shaped hexameric helicases and translocases support essential DNA, RNA, and protein-dependent transactions in all cells and many viruses. How such systems coordinate ATPase activity between multiple subunits to power conformational changes that drive the engagement and movement of client substrates is a fundamental question. Using the E. coli Rho transcription termination factor as a model system, we have employed solution and crystallographic structural methods to delineate the range of conformational changes that accompany distinct substrate and nucleotide cofactor binding events. SAXS data show that Rho preferentially adopts an open-ring state in solution, and that RNA and ATP are both required to cooperatively promote ring closure. Multiple closed-ring structures with different RNA substrates and nucleotide occupancies capture distinct catalytic intermediates accessed during translocation. Our data reveal how RNA-induced ring closure templates a sequential ATP-hydrolysis mechanism, provide a molecular rationale for how the Rho ATPase domains distinguishes between distinct RNA sequences, and establish the first structural snapshots of substepping events in a hexameric helicase/translocase.SIGNIFICANCEHexameric, ring-shaped translocases are molecular motors that convert the chemical energy of ATP hydrolysis into the physical movement of protein and nucleic acid substrates. Structural studies of several distinct hexameric translocases have provided insights into how substrates are loaded and translocated; however, the range of structural changes required for coupling ATP turnover to a full cycle of substrate loading and translocation has not been visualized for any one system. Here, we combine low-and high-resolution structural studies of the Rho helicase, defining for the first time the ensemble of conformational transitions required both for substrate loading in solution and for substrate movement by a processive hexameric translocase.

Molecular recognition with convergent functional groups. VI. Synthetic and structural studies with a model receptor for nucleic acid components

1989 ◽  
Vol 111 (3) ◽  
pp. 1082-1090 ◽  
Author(s):  
Ben Askew ◽  
Pablo Ballester ◽  
Chris Buhr ◽  
Kyu Sung Jeong ◽  
Sharon Jones ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Molecular Recognition ◽  
Functional Groups ◽  
Structural Studies
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