2014
DOI: 10.1021/ac4036636
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Interplay of Binding Stoichiometry and Recognition Specificity for the Interaction of MBD2b Protein and Methylated DNA Revealed by Affinity Capillary Electrophoresis Coupled with Laser-Induced Fluorescence Analysis

Abstract: ABSTRACT:The methyl-CpG binding domain (MBD) family proteins can specifically bind methylated DNA sequences and thereby mediate gene transcription. In this study, we used neutral capillary electrophoresis coupled with laser-induced fluorescence to investigate the interactions of DNA and MBD2b, a model MBD family protein with the highest affinity. For this purpose, we synthesized 13 double-stranded oligonucleotides of varying length (20 bp to 80 bp) and of varying methylation density. The sequences of these oli… Show more

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Cited by 17 publications
(19 citation statements)
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“…Many different assay schemes have been demonstrated for detecting and quantifying affinity interactions by MCE. [102104] Noncovalent interactions assayed by CE include protein-nucleic acid [9,103,105], protein-peptide [70,68,106], protein-protein [3,107110], nucleic acid-small molecule [111,112] and protein-small molecule interactions [113117]. Of these, a “mix and separate” approach, sometimes called affinity probe CE [118], is perhaps the most amenable for high throughput screening.…”
Section: Targetsmentioning
confidence: 99%
See 1 more Smart Citation
“…Many different assay schemes have been demonstrated for detecting and quantifying affinity interactions by MCE. [102104] Noncovalent interactions assayed by CE include protein-nucleic acid [9,103,105], protein-peptide [70,68,106], protein-protein [3,107110], nucleic acid-small molecule [111,112] and protein-small molecule interactions [113117]. Of these, a “mix and separate” approach, sometimes called affinity probe CE [118], is perhaps the most amenable for high throughput screening.…”
Section: Targetsmentioning
confidence: 99%
“…[9,104,105] In one example, the protein-nucleic acid interaction between human immunodeficiency virus 1 transactivator of transcription (Tat) and transactivation-responsive RNA (TAR) was studied using a commercial microchip platform. Inhibition of the Tat-TAR complex was demonstrated using a known inhibitor and dose dependent inhibition of the Tat-TAR complex peak was observed, suggesting the potential application of this platform to screening of affinity complexes.…”
Section: Targetsmentioning
confidence: 99%
“…Zou et al used ACE coupled with LIF to study binding stoichiometry and specific recognition between future science group Recent advances in ACE for binding studies Review methyl CPG-binding domain (MBD2b) protein and methylated DNA an interaction which is involved in gene transcription [46].…”
Section: Protein-dna Interactionsmentioning
confidence: 99%
“…Current-to-voltage converter was applied to convert the signal output from photomultiplier tube to voltage signal. The obtained electropherograms are shown in Figure 3 [46].…”
Section: Coupling With Fluorescence Detectormentioning
confidence: 99%
“…Zou et al. found that the DNA‐methyl‐CpG‐binding domain protein (MBD2b‐DNA) complex is a DNA length‐dependent binding stoichiometry and improved the determination of DNA methylation based on MBD proteins .…”
Section: Introductionmentioning
confidence: 99%