The web interface enables binding sites detection, virtual screening hits identification, and drug targets prediction in an interactive manner through a seamless interface to all adapted packages (e.g., Cavity, PocketV.2, PharmMapper, SHAFTS).
Here, we interrogate for the first time the low limits of fragment binding detection using a successfully targeted PPI as a model system.
However, a more general and simpler approach would be to increase the sensitivity of binding detection in a fragment screen.
The instrumental setup described by de Vlieger et al. [ 20] was used for the analysis, with one enzyme binding detection line and the following modifications (cf Fig. 1).
Enzyme binding detection was achieved with a competitive binding assay based on fluorescence enhancement which has a simple principle, is inexpensive, and is easy to interpret.
For this study, we used a modified version of the gp41 MPER peptide to enhance solubitiy and facilitate binding detection (EQELLELDKWASLGGGGSGGWNWFDITKWLWYIKKKKGSKKK).
Hence, the temperatures of the HPLC separation step and the enzyme binding detection were controlled separately.
Post-column, the flow was split 1 9 (Fig. 1, 3), sending 100 μL/min to high-resolution mass spectrometry (HR-MS) detection and 13 μL/min to the enzyme binding detection.
Especially, sensing systems using localized SPR (LSPR) have received significant research attention in recent years as a result of their potential for use as highly sensitive, simple, and label-free bio/chemical binding detection devices [4 6].
High-resolution mass spectrometry gives structural information to identify small molecules while an online enzyme binding detection method provides data on p38α binding.
