Chemical library screening for WNK signalling inhibitors using fluorescence correlation spectroscopy

2013 ◽  
Vol 455 (3) ◽  
pp. 339-345 ◽  
Author(s):  
Takayasu Mori ◽  
Eriko Kikuchi ◽  
Yuko Watanabe ◽  
Shinya Fujii ◽  
Mari Ishigami-Yuasa ◽  
...  

To discover WNK–OSR1/SPAK signalling inhibitors, we generated a new high-throughput system using fluorescent correlation spectroscopy capable of screening compounds that disrupt the binding of two molecules. We finally identified two novel and promising compounds for WNK–OSR1/SPAK signalling inhibition.


2007 ◽  
Vol 17 (10) ◽  
pp. 2904-2907 ◽  
Author(s):  
Yoichiro Hoshina ◽  
Yoshifumi Yamada ◽  
Hiroshi Tanaka ◽  
Takayuki Doi ◽  
Takashi Takahashi






2020 ◽  
Vol 412 (11) ◽  
pp. 2589-2597 ◽  
Author(s):  
Xu Fu ◽  
Yongwook Song ◽  
Abdullah Masud ◽  
Kanthi Nuti ◽  
Jason E. DeRouchey ◽  
...  


1999 ◽  
Vol 4 (6) ◽  
pp. 335-353 ◽  
Author(s):  
Keith J. Moore ◽  
Sandra Turconi ◽  
Stephen Ashman ◽  
Martin Ruediger ◽  
Ulrich Haupts ◽  
...  

Fluorescence assay technologies used for miniaturized high throughput screening are broadly divided into two classes. Macroscopic fluorescence techniques (encompassing conventional fluorescence intensity, anisotropy [also often referred to as fluorescence polarization] and energy transfer) monitor the assay volume- and time-averaged fluorescence output from the ensemble of emitting fluorophores. In contrast, single-molecule detection (SMD) techniques and related approaches, such as fluorescence correlation spectroscopy (FCS), stochastically sample the fluorescence properties of individual constituent molecules and only then average many such detection events to define the properties of the assay system as a whole. Analysis of single molecular events is accomplished using confocal optics with an illumination/detection volume of -1 fl (10-15 L) such that the signal is insensitive to miniaturization of HTS assays to 1 A1 or below. In this report we demonstrate the general applicability of one SMD technique (FCS) to assay configuration for target classes typically encountered in HTS and confirm the equivalence of the rate/equilibrium constants determined by FCS and by macroscopic techniques. Advantages and limitations of the current FCS technology, as applied here, and potential solutions, particularly involving alternative SMD detection techniques, are also discussed.



2009 ◽  
Vol 18 (5) ◽  
pp. 1115-1120 ◽  
Author(s):  
Toshihiko Sugiki ◽  
Chie Yoshiura ◽  
Yutaka Kofuku ◽  
Takumi Ueda ◽  
Ichio Shimada ◽  
...  


2015 ◽  
Vol 33 (4) ◽  
pp. 384-389 ◽  
Author(s):  
Malte Wachsmuth ◽  
Christian Conrad ◽  
Jutta Bulkescher ◽  
Birgit Koch ◽  
Robert Mahen ◽  
...  


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