Dixon plots of em K /em i data shown in Fig. of the enzyme. Analysis of 4 via these methods proved problematic due to the presence of an intrinsic fluorescence in the compound. This was a particular problem with the enzymes, as competitive inhibitors, with micromolar inhibitory activity (S3 Fig.). Even though identified compounds inhibit both position of the 2-phenyl ring) were unable to coordinate the zinc ion(s) of either enzyme (S4 Fig. and S5 Fig.). In existence cycle. Supporting Info S1 FigPrimary display of MMV400 using a multiplex aminopeptidase assay. Enzyme activity that was reduced in comparison to control wells are Engeletin indicated by R and compounds that experienced no effect on activity are demonstrated as UC for unchanged. Compounds highlighted in gray are drug-like and the remainder are probe-like. (PDF) Click here for more data file.(99K, pdf) S2 FigSecondary display of em Pf /em A-M1 and em Pf /em A-M17 against 24 initial screen hits. Enzyme activity in the presence of 100 M compound (MMV# demonstrated) was compared to activity of the enzyme in the absence of any inhibitor (-). An inhibitor control using Bestatin was included and no neutral aminopeptidase activity is definitely detectable in the presence of 100 M Bestatin. A dashed collection is shown to indicate the when the activity of either enzyme was reduced by 90% or more. (PDF) Click here for more data file.(94K, pdf) S3 FigInhibitory properties of MMV666023 and MMV020750. (A) Enzyme activity in the presence of increasing inhibitor concentration. Numbers demonstrated on curves are inhibitor concentration in M. (B) Dixon storyline for calculation of em K /em i where S1 and S2 are two different substrate concentrations ( em K /em M of enzyme). (PDF) Click here for more data file.(270K, pdf) S4 FigDixon Rabbit polyclonal to ATF6A plots and docking for MMV020750 derivatives with em Pf /em A-M1. Dixon plots of em K /em i Engeletin data demonstrated in Fig. 5 ( em K Engeletin /em i defined as point of intersection and indicated by dotted collection). Two different substrate concentrations are demonstrated (solid circles and squares). Outliers not included in Engeletin linear regression are demonstrated as hollow squares or circles. 3D molecular docking diagrams demonstrated with Engeletin carbon atoms of em Pf /em A-M17 residues and the inhibitor are coloured in light and dark gray, respectively. Zinc ions are demonstrated as spheres. Related 2D molecular docking representations demonstrated on right hand panel. (PDF) Click here for more data file.(2.2M, pdf) S5 FigDixon plots and docking for MMV020750 derivatives with em Pf /em A-M17. Dixon plots of em K /em i data demonstrated in Fig. 5 ( em K /em i defined as point of intersection and indicated by dotted collection except in 4 where a grey shaded area defines em K /em i range). Two different substrate concentrations are demonstrated (solid circles and squares). Outliers not included in linear regression are demonstrated as hollow squares or circles. 3D molecular docking diagrams demonstrated with carbon atoms of em Pf /em A-M17 residues and the inhibitor are coloured in light and dark gray, respectively. Zinc ions are demonstrated as spheres. Related 2D molecular docking representations demonstrated on right hand panel. (PDF) Click here for more data file.(2.8M, pdf) S1 MethodsChemistry Experimental. (PDF) Click here for more data file.(124K, pdf) Funding Statement The work was supported by an Australian National Health and Medical Study Council Project Give to SM and PJS (1063786). SM is an Australian Study Council Long term Fellow (Feet100100690). The funders experienced no part in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Data Availability All relevant data are within the paper and its Supporting Information documents..