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As an initial step to define molecular mechanism of TB10-dependent apoptotic process in ovarian tumor cells, we searched a human ovary cDNA library for a novel TB10 binding protein using a yeast two-hybrid system.
Shortly after the identification of ASK1, ASK2 (also designated as MAP3K6) was identified as an ASK1 binding protein using a yeast two-hybrid screen [ 37].
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In order to further assess the role of MAD2B in cell cycle control, we set out to identify binding proteins using a yeast two-hybrid interaction trap (see Materials and methods).
AATYK2/cprk was isolated as a p35-binding protein using a yeast two-hybrid system [5], as was the case for AATYK1 [15].
When we screened the Ci-binding proteins using a yeast two-hybrid assay, we have also identified importin α3, in addition to Rdx (see Materials and methods).
Subsequently, we confirmed properties of this protein as an I2 binding protein using biochemical and pharmacological techniques.
Previous reports have demonstrated small molecule binding to protein using a high density sensor surface and suggest that aptamer immobilization at higher densities may support a more sensitive and stable method for measuring small molecule aptamer binding.
Finally, we describe a method for visualizing dynamic assembly and disassembly of single actin filaments and fluorescently labeled actin binding proteins using total internal reflection fluorescence (TIRF) microscopy.
A recent report characterizing EGFR as a DNA-binding protein using unbiased approaches [ 95] further supports the notion that the nuclear EGFR family plays a role in transcriptional regulation.
In this process, ligands were automatically docked into the binding site of the protein using a ProtoMol-based approach [37] with a patented search engine and an empirical scoring function.
Using RNA polymerase as a model system, our laboratory has shown that it is possible to measure the apparent (or effective) target size of a DNA-binding protein using DNA curtains to visualize how RNAP searches for promoters in real time.
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