Chemoenzymatic Synthesis and High-Throughput Screening of an Aminoglycoside−Polyamine Library:  Identification of High-Affinity Displacers and DNA-Binding Ligands

2004 ◽  
Vol 126 (39) ◽  
pp. 12306-12315 ◽  
Author(s):  
Kaushal Rege ◽  
Shanghui Hu ◽  
James A. Moore ◽  
Jonathan S. Dordick ◽  
Steven M. Cramer
Keyword(s):  
Dna Binding ◽  
High Throughput ◽  
High Throughput Screening ◽  
Chemoenzymatic Synthesis ◽  
High Affinity

Chemoenzymatic synthesis of Park’s nucleotide: toward the development of high-throughput screening for MraY inhibitors

2007 ◽  
Vol 48 (5) ◽  
pp. 799-803 ◽  
Author(s):  
Michio Kurosu ◽  
Sebabrata Mahapatra ◽  
Prabagaran Narayanasamy ◽  
Dean C. Crick
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Chemoenzymatic Synthesis

scholarly journals Identification of HMGA2 inhibitors by AlphaScreen-based ultra-high-throughput screening assays

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Linjia Su ◽  
Nadezda Bryan ◽  
Sabrina Battista ◽  
Juliano Freitas ◽  
Alyssa Garabedian ◽  
...  
Keyword(s):  
Dna Binding ◽  
High Throughput ◽  
Dna Sequences ◽  
High Throughput Screening ◽  
Dna Interactions ◽  
High Mobility Group Protein ◽  
Screening Assay ◽  
Binding Motifs ◽  
Dna Binding Motifs ◽  
High Throughput Screening Assay

Abstract The mammalian high mobility group protein AT-hook 2 (HMGA2) is a multi-functional DNA-binding protein that plays important roles in tumorigenesis and adipogenesis. Previous results showed that HMGA2 is a potential therapeutic target of anticancer and anti-obesity drugs by inhibiting its DNA-binding activities. Here we report the development of a miniaturized, automated AlphaScreen ultra-high-throughput screening assay to identify inhibitors targeting HMGA2-DNA interactions. After screening the LOPAC1280 compound library, we identified several compounds that strongly inhibit HMGA2-DNA interactions including suramin, a century-old, negatively charged antiparasitic drug. Our results show that the inhibition is likely through suramin binding to the “AT-hook” DNA-binding motifs and therefore preventing HMGA2 from binding to the minor groove of AT-rich DNA sequences. Since HMGA1 proteins also carry multiple “AT-hook” DNA-binding motifs, suramin is expected to inhibit HMGA1-DNA interactions as well. Biochemical and biophysical studies show that charge-charge interactions and hydrogen bonding between the suramin sulfonated groups and Arg/Lys residues play critical roles in the binding of suramin to the “AT-hook” DNA-binding motifs. Furthermore, our results suggest that HMGA2 may be one of suramin’s cellular targets.

scholarly journals Combinatorial Chemoenzymatic Synthesis and High-Throughput Screening of Sialosides

2008 ◽  
Vol 3 (9) ◽  
pp. 567-576 ◽  
Author(s):  
Harshal A. Chokhawala ◽  
Shengshu Huang ◽  
Kam Lau ◽  
Hai Yu ◽  
Jiansong Cheng ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Chemoenzymatic Synthesis

scholarly journals Early Identification of False Positives in High-Throughput Screening for Activators of p53-DNA Interaction

2006 ◽  
Vol 11 (4) ◽  
pp. 341-350 ◽  
Author(s):  
Julian Wölcke ◽  
Nicholas Hunt ◽  
Joern Jungmann ◽  
Dirk Ullmann
Keyword(s):  
Dna Binding ◽  
High Throughput ◽  
High Throughput Screening ◽  
Fold Increase ◽  
Dna Interaction ◽  
False Positives ◽  
Correlation Spectroscopy ◽  
Detection Methods ◽  
Distribution Analysis ◽  
Process Step

Naturally occurring mutant forms of p53 are deficient for specific DNA binding. However, their specific DNA binding can be reactivated. The search for small molecules that reactivate latent p53 is considered to be a cornerstone in cancer therapy. The authors describe a new homogeneous fluorescent assay approach for the characterization of binding affinities of human wild-type latent and activated p53 using DNA*spec(26), with and without the addition of the antibody PAb421, respectively, and fluorescence correlation spectroscopy (FCS)/2-dimensional fluorescence-intensity distribution analysis anisotropy as the detection methods. FCS was compared with 2D-FIDA anisotropy, and a very good correlation of the results with both readouts was observed (KDs for nonspecific DNA binding of 24.4 ± 3.5 nM with 2D-FIDA anisotropy and of 29.5 ± 5.5 nM with FCS). The presence of poly(dI-dC) led to a 10-fold increase of binding affinity (KD of 3.3 ± 0.5 nM in the presence of PAb421). 2D-FIDA anisotropy was demonstrated to be the most accurate readout; hence, this detection technology was selected for a 25,000 compound member high-throughput screening (HTS) campaign. The hits obtained were qualified using a novel data evaluation algorithm that identifies false positives and moreover assesses the validity of true hits in the presence of the deteriorating artifact. This process step is of utmost importance for decreasing the attrition in fluorescence-based HTS.

High-Throughput Screening for High Affinity Antibodies

2009 ◽  
Vol 14 (5) ◽  
pp. 303-307 ◽  
Author(s):  
Simon Tickle ◽  
Ralph Adams ◽  
Derek Brown ◽  
Meryn Griffiths ◽  
Daniel Lightwood ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
High Affinity
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