Nonequilibrium Capillary Electrophoresis of Equilibrium Mixtures − A Single Experiment Reveals Equilibrium and Kinetic Parameters of Protein−DNA Interactions

2002 ◽  
Vol 124 (46) ◽  
pp. 13674-13675 ◽  
Author(s):  
Maxim Berezovski ◽  
Sergey N. Krylov
Keyword(s):  
Capillary Electrophoresis ◽  
Kinetic Parameters ◽  
Dna Interactions ◽  
Single Experiment ◽  
Protein Dna Interactions

scholarly journals Analysis of protein-DNA interactions in chromatin by UV induced cross-linking and mass spectrometry

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Alexandra Stützer ◽  
Luisa M. Welp ◽  
Monika Raabe ◽  
Timo Sachsenberg ◽  
Christin Kappert ◽  
...  
Keyword(s):  
Uv Light ◽  
Amino Acid Level ◽  
Dna Interaction ◽  
Cross Linking ◽  
Dna Interactions ◽  
Single Experiment ◽  
Protein Dna Interactions ◽  
Nucleosome Binding ◽  
Binding Interfaces

Abstract Protein–DNA interactions are key to the functionality and stability of the genome. Identification and mapping of protein–DNA interaction interfaces and sites is crucial for understanding DNA-dependent processes. Here, we present a workflow that allows mass spectrometric (MS) identification of proteins in direct contact with DNA in reconstituted and native chromatin after cross-linking by ultraviolet (UV) light. Our approach enables the determination of contact interfaces at amino-acid level. With the example of chromatin-associated protein SCML2 we show that our technique allows differentiation of nucleosome-binding interfaces in distinct states. By UV cross-linking of isolated nuclei we determined the cross-linking sites of several factors including chromatin-modifying enzymes, demonstrating that our workflow is not restricted to reconstituted materials. As our approach can distinguish between protein–RNA and DNA interactions in one single experiment, we project that it will be possible to obtain insights into chromatin and its regulation in the future.

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