Analysis of binding specificity of disulfide bonded dimeric lambda-Cro V55C protein with generic hexamer oligonucleotide microchipстатья
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Дата последнего поиска статьи во внешних источниках: 4 октября 2013 г.
Аннотация:Binding specificity of mutant V55C disulfide bonded dimeric lambda-Cro protein
(CroVC) to double-stranded DNA (dsDNA) was studied using generic hexamer
oligonucleotide microchip. The curves of dissociation of hybridized DNA in the
presence and absence of CroVC were converted into the effective discriminant
constants to assess the relevant thermodynamic equilibrium binding constants for
dsDNA-protein complexes. Then, tiling of longer oligonucleotides with shorter
oligomers was used to search for sequence motifs with the highest binding
specificity similarly to sequencing by hybridization. The comparison of the
deduced sequences with the known natural operator half-sites demonstrated the
principal ability to discern and reconstruct the major parts of 7-mer motifs
corresponding to the strongest binding of CroVC subunits. Our results show the
applicability of generic microchips to the analysis of binding specificity in the
case of multi-subunit DNA-binding proteins.