~10nM - 10µM
||Budding yeast Saccharomyces cerevisiae
||Fordyce PM et al., De novo identification and biophysical characterization of transcription-factor binding sites with microfluidic affinity analysis. Nat Biotechnol. 2010 Sep28(9):970-5. doi: 10.1038/nbt.1675. p.972 right column top paragraphPubMed ID20802496
|| Maerkl, S.J. & Quake, S.R. A systems approach to measuring the binding energy landscapes of transcription factors. Science 315, 233–237 (2007).PubMed ID17218526
||Reference abstract: "[Investigators] present a microfluidics-based approach for de novo discovery and quantitative biophysical characterization of DNA target sequences. [They] validated [their] technique by measuring sequence preferences for 28 Saccharomyces cerevisiae transcription factors with a variety of DNA-binding domains, including several that have proven difficult to study by other techniques." Primary source abstract: "Measuring affinities of molecular interactions in high-throughput format remains problematic, especially for transient and low-affinity interactions. [Investigators] describe a high-throughput microfluidic platform that measures such properties on the basis of mechanical trapping of molecular interactions."
||P.972 right column top paragraph: "The range of Kd values calculated here for Pho4p and Cbf1p agree with those measured in previous studies (~10 nM–10 μM) [primary source], validating [investigators'] approach." PMID 15111622 abstract:"Cbf1p is a basic-helix-loop-helix-zipper protein of Saccharomyces cerevisiae required for the function of centromeres and MET gene promoters, where it binds DNA via the consensus core motif CACRTG (R = A or G)." Pho4p is a transcriptional activator of the PHO regulon.