Journal: Marine Drugs

Article Title: Structure–Activity Relationship Studies Using Natural and Synthetic Okadaic Acid/Dinophysistoxin Toxins

doi: 10.3390/md14110207

Figure Lengend Snippet: Effect of various DSP toxins on protein phosphatase activity. The activities of protein phosphatases ( A ) PP2a, ( B ) PP1, and ( C ) PP5 were assessed in the presence of various concentrations of natural OA, natural DTX1, natural DTX2, synthetic DTX2, or synthetic 2- epi -DTX2. All data (mean ± SE; n = 3 or 4) were normalized to the control. Three-parameter, variable slope, non-linear dose-response analysis was performed and IC 50 and 95% confidence interval values were calculated . Note: Some of these data have been published previously in Pang et al. .

Article Snippet: Briefly, the assay tests the ability of toxins to inhibit the activity of purified PP2A (EMD Millipore Corporation, Darmstadt, Germany, cat. #14-111; human red blood cells), purified PP1 (EMD Millipore Corporation, Darmstadt, Germany, cat. #14-110; rabbit skeletal muscle), or purified PP5 (EMD Millipore Corporation, Darmstadt, Germany, cat. #14-778; recombinant enzyme expressed in E. coli cells) against the fluorimetric substrate, 6,8-difluro-4-methyl umbelliferyl phosphate (DiFMUP; 50 μM final) (Invitrogen ThermoFisher Scientific, Waltham, MA, USA, cat. #D6567).

Techniques: Activity Assay, Control

Journal: Marine Drugs

Article Title: Structure–Activity Relationship Studies Using Natural and Synthetic Okadaic Acid/Dinophysistoxin Toxins

doi: 10.3390/md14110207

Figure Lengend Snippet: Calculated IC 50 values and relative potencies for natural OA, DTX1, and DTX2, and synthetic DTX2, and 2 -epi -DTX2, based on PP2a, PP1, and PP5 inhibition.

Article Snippet: Briefly, the assay tests the ability of toxins to inhibit the activity of purified PP2A (EMD Millipore Corporation, Darmstadt, Germany, cat. #14-111; human red blood cells), purified PP1 (EMD Millipore Corporation, Darmstadt, Germany, cat. #14-110; rabbit skeletal muscle), or purified PP5 (EMD Millipore Corporation, Darmstadt, Germany, cat. #14-778; recombinant enzyme expressed in E. coli cells) against the fluorimetric substrate, 6,8-difluro-4-methyl umbelliferyl phosphate (DiFMUP; 50 μM final) (Invitrogen ThermoFisher Scientific, Waltham, MA, USA, cat. #D6567).

Techniques: Inhibition

Journal: The Journal of Biological Chemistry

Article Title: The Molecular Chaperone Hsp70 Activates Protein Phosphatase 5 (PP5) by Binding the Tetratricopeptide Repeat (TPR) Domain *

doi: 10.1074/jbc.M113.519421

Figure Lengend Snippet: TABLE 2

Article Snippet: The two spectra have the same intrinsic signal to noise ratio ((254/169) 2 = 2.25). p-Nitrophenyl Phosphate Assay Purified PP5, Hsp70, and Hsp90 were immobilized in 4HBX 96-well plates (Thermo Scientific) and diluted with ELISA buffer (BioLegend).

Techniques: