Covalent Binding of Modified Nucleic Acids to Proteins as a Method for Investigation of Specific Protein—Nucleic Acid Interactions

ChemInform ◽  
2005 ◽  
Vol 36 (28) ◽  
Author(s):  
T. S. Zatsepin ◽  
N. G. Dolinnaya ◽  
E. A. Kubareva ◽  
M. G. Ivanovskaya ◽  
V. G. Metelev ◽  
...  
2005 ◽  
Vol 74 (1) ◽  
pp. 77-95 ◽  
Author(s):  
Timofei S Zatsepin ◽  
N G Dolinnaya ◽  
Elena A Kubareva ◽  
Marina G Ivanovskaya ◽  
V G Metelev ◽  
...  

Author(s):  
Stephen D. Jett

The electrophoresis gel mobility shift assay is a popular method for the study of protein-nucleic acid interactions. The binding of proteins to DNA is characterized by a reduction in the electrophoretic mobility of the nucleic acid. Binding affinity, stoichiometry, and kinetics can be obtained from such assays; however, it is often desirable to image the various species in the gel bands using TEM. Present methods for isolation of nucleoproteins from gel bands are inefficient and often destroy the native structure of the complexes. We have developed a technique, called “snapshot blotting,” by which nucleic acids and nucleoprotein complexes in electrophoresis gels can be electrophoretically transferred directly onto carbon-coated grids for TEM imaging.


Author(s):  
Pamela S. Hicks ◽  
Christopher L. Andrews ◽  
David G. Bear

It is common for sing1e-stranded nucleic acid finding proteins to bind polynucleotides cooperatively. The parameter ω is used to describe the component of the association binding constant that is due to cooperative interactions between protein molecules on the polynucleotide lattice; ω is 1 for noncooperative binding, while ω can be as high as 103-104 for highly cooperative proteins such as bacteriophage T4 gene 32 protein. In the past, to has been determined by computer fitting of spectroscopic titration data. It has been suggested that electrom microscopy would provide a more direct method for estimation of the cooperativity parameter. An equation can be derived that relates the experimentally obtained average protein cluster size (at a specific protein/RNA saturation ratio) to ω. The only other parameter required for the calculation is the RNA binding site size obtained from stoichiometric titration measurements.


Biochemistry ◽  
1980 ◽  
Vol 19 (15) ◽  
pp. 3516-3522 ◽  
Author(s):  
Timothy M. Lohman ◽  
C. Glen Wensley ◽  
Jeffrey Cina ◽  
Richard R. Burgess ◽  
M. Thomas Record

1998 ◽  
Vol 8 (1) ◽  
pp. 9-10 ◽  
Author(s):  
PhilipE Bourne ◽  
Judith Murray-Rust ◽  
JeremyH Lakey

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