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Ultrafast differential flexibility of Cro-protein binding domains of two operator DNAs with different sequences

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Title Ultrafast differential flexibility of Cro-protein
binding domains of two operator DNAs with
different sequences
 
Creator Choudhury, Susobhan
Ghosh, Basusree
Singh, Priya
Ghosh, Raka
Roy, Siddhartha
Pal, Samir Kumar
 
Subject Structural Biology & Bioinformatics
 
Description The nature of the interface of specific protein–DNA complexes has attracted immense interest in contemporary
molecular biology. Although extensive studies on the role of flexibility of DNA in the specific interaction
in the genetic regulatory activity of lambda Cro (Cro-protein) have been performed, the exploration of
quantitative features remains deficient. In this study, we have mutated (site directed mutagenesis: SDM)
Cro-protein at the 37th position with a cysteine residue (G37C) retaining the functional integrity of the
protein and labelled the cysteine residue, which is close to the interface, with a fluorescent probe
(AEDANS), for the investigation of its interface with operator DNAs (OR3 and OR2). We have employed
picosecond resolved polarization gated fluorescence spectroscopy and the well known strategy of
solvation dynamics for the exploration of physical motions of the fluorescent probes and associated
environments, respectively. Even though this particular probe on the protein (AEDANS) shows marginal
changes in its structural flexibility upon interaction with the DNAs, a non-covalent DNA bound probe
(DAPI), which binds to the minor groove, shows a major differential alteration in the dynamical flexibility
in the OR3–Cro complex when compared to that of the OR2 complex with the Cro-protein. We attempt
to correlate the observed significant structural fluctuation of the Cro-protein binding domain of OR3 for
the specificity of the protein to the operator DNA
 
Date 2016
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://www.eprints.iicb.res.in/2546/1/Chem._Phys.%2C_2016%2C18%2C_17983%2D17990.pdf
Choudhury, Susobhan and Ghosh, Basusree and Singh, Priya and Ghosh, Raka and Roy, Siddhartha and Pal, Samir Kumar (2016) Ultrafast differential flexibility of Cro-protein binding domains of two operator DNAs with different sequences. Phys. Chem. Chem. Phys, 18. pp. 17983-17990.
 
Relation http://www.eprints.iicb.res.in/2546/