scholarly journals Development of high-throughput screening assays for profiling snake venom phospholipase A2 activity after chromatographic fractionation

Toxicon ◽  
2020 ◽  
Vol 184 ◽  
pp. 28-38 ◽  
Author(s):  
Kristina B.M. Still ◽  
Julien Slagboom ◽  
Sarah Kidwai ◽  
Chunfang Xie ◽  
Yumei Zhao ◽  
...  
Keyword(s):  
Phospholipase A2 ◽  
Snake Venom ◽  
High Throughput ◽  
High Throughput Screening ◽  
Chromatographic Fractionation ◽  
Snake Venom Phospholipase ◽  
Screening Assays ◽  
Phospholipase A2 Activity

scholarly journals Development of high-throughput screening assays for profiling snake venom Phospholipase A2 activity after high-resolution chromatographic fractionation

2020 ◽  
Author(s):  
Kristina B.M. Still ◽  
Julien Slagboom ◽  
Sarah Kidwai ◽  
Chunfang Xie ◽  
Bastiaan Eisses ◽  
...  
Keyword(s):  
Fatty Acid ◽  
High Resolution ◽  
High Throughput ◽  
High Throughput Screening ◽  
Proinflammatory Mediators ◽  
Ph Indicators ◽  
Snake Venom Phospholipase ◽  
Screening Assays ◽  
Animal Venoms ◽  
Phospholipase A2 Activity

AbstractMany organisms, ranging from plants to mammals, contain phospholipase A2 enzymes (PLA2s), which catalyze the production of lysophospholipids and fatty acid proinflammatory mediators. PLA2s are also common constituents of animal venoms, including bees, scorpions and snakes, and they cause a wide variety of toxic effects including neuro-, myo-, cyto-, and cardio-toxicity, anticoagulation and edema. The aim of this study was to develop a generic method for profiling enzymatically active PLA2s in snake venoms after chromatographic separation. For this, low-volume high-throughput assays for assessment of enzymatic PLA2 activity were evaluated and optimized. Subsequently, the assays were incorporated into a nanofractionation platform that combines high-resolution fractionation of crude venoms by liquid chromatography (LC) with bioassaying in 384-well plate format, and parallel mass spectrometric (MS) detection for toxin identification. The miniaturized assays developed are based on absorbance or fluorescence detection (respectively, using cresol red or fluorescein as pH indicators) to monitor the pH drop associated with free fatty acid formation by enzymatically active PLA2s. The methodology was demonstrated for assessment of PLA2 activity profiles of venoms from the snake species Bothrops asper, Echis carinatus, Echis coloratus, Echis ocellatus, Oxyuranus scutellatus and Daboia russelii russelii.

scholarly journals Design of High-Throughput Screening Assays and Identification of a SUMO1-Specific Small Molecule Chemotype Targeting the SUMO-Interacting Motif-Binding Surface

2015 ◽  
Vol 17 (4) ◽  
pp. 239-246 ◽  
Author(s):  
Aileen Y. Alontaga ◽  
Yifei Li ◽  
Chih-Hong Chen ◽  
Chen-Ting Ma ◽  
Siobhan Malany ◽  
...  
Keyword(s):  
Small Molecule ◽  
High Throughput ◽  
High Throughput Screening ◽  
Binding Surface ◽  
Screening Assays

scholarly journals Development of high throughput screening assays and pilot screen for inhibitors of metalloproteases meprin α and β

Biopolymers ◽  
2014 ◽  
Vol 102 (5) ◽  
pp. 396-406 ◽  
Author(s):  
Franck Madoux ◽  
Claudia Tredup ◽  
Timothy P. Spicer ◽  
Louis Scampavia ◽  
Peter S. Chase ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Screening Assays
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