Dataset of cathepsin L-like CP inhibition of Naegleria fowleri and Acanthamoeba castellanii by ppTvCP4r from Trichomonas vaginalis

The recombinant TvCP4 prepro region (ppTvCP4r) acts as an exogenous inhibitor of cathepsin L-like CPs from Trichomonas vaginalis (Cárdenas-Guerra et al., 2015 [1]). Here, we present the dataset of the trichomonad ppTvCP4r inhibitory effect against the CP proteolytic activities from other microorganisms, such as Naegleria fowleri and Acanthamoeba castellanii free-living amoeba. The proteolytic activity inhibition of total crude extracts (TCEs) of N. fowleri and A. castellanii was determined and recorded using a fluorogenic substrate specific for cathepsin L CPs without or with a ppTvCP4r treatment at different concentrations and pH.


a b s t r a c t
The recombinant TvCP4 prepro region (ppTvCP4r) acts as an exogenous inhibitor of cathepsin L-like CPs from Trichomonas vaginalis (Cárdenas-Guerra et al., 2015 [1]). Here, we present the dataset of the trichomonad ppTvCP4r inhibitory effect against the CP proteolytic activities from other microorganisms, such as Naegleria fowleri and Acanthamoeba castellanii free-living amoeba. The proteolytic activity inhibition of total crude extracts (TCEs) of N. fowleri and A. castellanii was determined and recorded using a fluorogenic substrate specific for cathepsin L CPs without or with a ppTvCP4r treatment at different concentrations and pH. &

Value of the data
The data show the ability of a fluorogenic substrate to detect low levels of CP proteolytic activity not detectable by zymography.
The data shows the potent enzyme inhibitory action of the recombinant prepro region of cathepsin L-like CP from Trichomonas vaginalis (ppTvCP4r) on CPs from free-living amoeba Naegleria fowleri and Acanthamoeba castellanii.
The data shows the potential use of the ppTvCP4r inhibitor to help determine the potential role of CPs in the pathogenesis of this free-living amoeba that could open up its further potential use for drug targeting.
The data also shows the potential use of T. vaginalis ppTvCP4r against cathepsin L-like CPs from other organisms including human pathogens.

Data
The prepro regions of the cathepsin L-like cysteine proteinases (CPs) are also inhibitors of related peptidases, in which the selectivity correlates with the degree of similarity in the prepro region sequences of the target proteinases (Wiederanders et al., 2003 [2]; Yamamoto et al., 2002 [3]). The development of CP inhibitors has provided useful tools to study and to profile the cellular proteolytic activity, to identify their extra-lysosomal functions in cells and pathogen organisms and for potential application in medicine as candidates for antiparasitic chemotherapy, among other uses (Turk et al., 2002 [4]; Sajid and McKerrow, 2002 [5]). While the inhibitors developed may not necessarily prove useful as drugs because of the disadvantages that present such as bioavailability, low toxicity, and selectivity, they still have immense value as research tools in studying the biological function of targeted enzymes (Dubin, 2005 [6]). The dataset of this article provides information on the ppTvCP4r (trichomonad recombinant prepro region of TvCP4) inhibitory activity against cathepsin L-like proteases from Naegleria fowleri and Acanthamoeba castellanii free-living amoeba using a specific fluorogenic substrate ( Fig. 1 and Table 1). The concentration and time-dependent CP proteolytic activities at pH 5 and 7 in the absence and presence of ppTvCP4r have been recorded and presented.  Table 1). The error bars indicate the standard errors of the mean (SEM) of at least three independent experiments in triplicate. Significant differences (P o 0.001) between the results are marked with asterisks.

Enzyme inhibition assays
All enzyme inhibition assays were performed at 25°C for 120 s using a fluorogenic substrate (Z-Phe-Arg-AMC; Peptide Institute Inc., Osaka, Japan) specific for cathepsin L CPs. The reaction was initiated by the addition of the fluorogenic substrate into the reaction wells of a 96-well plate containing total crude extracts (TCEs). The increase in fluorescence intensity due to the release of aminomethyl coumarin (AMC) was measured using a Gemini EM Microplate Reader spectrofluorometer (SpectraMaxR Gemini EM; Molecular Devices, Sunnyvale, CA, USA) at 355 and 460 nm excitation and emission wavelengths, respectively. For all proteolytic activity inhibition assays, the kinetics were   Table 1. Statistically significant differences between the means were determined by analysis of variance (ANOVA) using Graph Pad Prism 5.0. The data were analyzed by one-way ANOVA using the Bonferroni method. All pairs of columns in Fig. 1 were compared (P o 0.001). The scores with statistically significant differences are indicated with asterisks in the figure. The corresponding P values are indicated in the figure legend.