A capillary electrophoresis method with dynamic pH junction stacking for the monitoring of XXXX sulfotransferase.

@entity15227 @entity4613 @entity29725 @entity9850 @entity8431 @entity45569 @entity74 @entity2786 @entity16038 @entity629 @entity597
UNASSIGNED: @entity8431 ( @entity8431 ) is a rare and severe @entity74 . Inhibition of @entity629 sulfotransferase (CST) has been proposed as a promising new therapeutic strategy for the treatment of @entity8431 . CST catalyzes the transfer of a @entity597 group from the coenzyme @entity16038 ( @entity16038 ) to @entity629 yielding @entity29725 and @entity45569 ( @entity2786 ). So far only a few weak CST inhibitors have been described. The goal of the present study was to establish a suitable assay for identifying and characterizing novel CST inhibitors. To this end, we developed and optimized a capillary electrophoresis (CE) based assay for monitoring the catalytic activity of CST by measuring the formation of PAP. A sample matrix consisting of 5mM @entity4613 buffer with about 0.0001% @entity9850 at pH 7.4 and a background electrolyte (BGE) containing 75mM @entity4613 buffer with 0.002% @entity9850 at pH 5.6 were utilized to achieve a stacking effect for PAP by dynamic pH junction. This led to a limit of detection for the enzymatic product PAP of 66.6nM. The CE method was sensitive, robust, and suitable for CST inhibitor screening, Ki value determination, and enzyme kinetic studies. Selected reference compounds were tested in order to validate the assay, including the substrates @entity629 and @entity15227 , and the inhibitor Congo Red. The newly developed CE method will be useful for the identification and development of novel CST inhibitors which are urgently needed for the treatment of @entity8431 .
