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Langmuir Monolayer as a Tool toward Visualization of a Specific DNA-Protein Complex

Brar, Loveleen K and Rajdev, Priya and Raychaudhuri, Arup K and Chatterji, Dipankar (2005) Langmuir Monolayer as a Tool toward Visualization of a Specific DNA-Protein Complex. In: Langmuir, 21 (23). pp. 10671-10675.

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Immobilization and imaging of protein molecules and protein-DNAcomplexes on a Langmuir-Blodgett (LB) substrate have been explored here. We have prepared a nickel-arachidate (NiA) monolayer and characterized it through pressure-area isotherm on a LB trough. Recombinant RNA polymerase from Escherichia coli, where the largest subunit was replaced with the same gene having a series of histidine amino acids at the C-terminus end of the protein, was immobilized over the Ni-arachidate monolayer through a Ni(II)-histidine interaction. A single molecule of RNA polymerase could be seen through intermittent-contact atomic force microscopy (AFM). Under the condition of the formation of the LB monolayer, the enzyme molecules were arrayed and transcriptionally active. Interestingly, they could pick up sequence specificDNAmolecules from the subphase in an oriented fashion.Onthe other hand, preformed RNA polymerase Ni(II)-arachidate monolayers bound DNA haphazardly when no surface pressure was employed.

Item Type: Journal Article
Publication: Langmuir
Publisher: American Chemical Society
Additional Information: The copyright belongs to American Chemical society.
Department/Centre: Division of Biological Sciences > Molecular Biophysics Unit
Division of Physical & Mathematical Sciences > Physics
Date Deposited: 22 Feb 2006
Last Modified: 19 Sep 2010 04:23
URI: http://eprints.iisc.ac.in/id/eprint/5528

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