unctions, glycosyltransferases are identified to be involved inside a multitude of biological processes, for instance cell ell communication, immune responses [4], cell signaling and epigenetic regulation of gene expression [7,8], and plant- and bacterial-cell wall biosynthesis [9,10]. As a corollary, the disruption of those biological processes as a consequence of abnormal glycosyltransferase CDK1 Inhibitor MedChemExpress activity or expression can possess a detrimental effect on thePublisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.Copyright: 2021 by the authors. Licensee MDPI, Basel, Switzerland. This short article is definitely an open access short article distributed under the terms and situations on the Inventive Commons Attribution (CC BY) license ( creativecommons.org/licenses/by/ 4.0/).Molecules 2021, 26, 6230. doi.org/10.3390/moleculesmdpi/journal/moleculesMolecules 2021, 26,two ofcell, major to really serious illnesses, like cancer, inflammation, and diabetes [11,12]. Glycosyltransferase inhibitors are becoming developed for the therapy of these diseases, as well as metabolic diseases, for instance Morbus Gaucher, a lysosomal storage disease characterized by an accumulation of glucocerebrosides in multiple organs because of dysfunctional downstream degradation machinery (glucocerebrosidase), causing detrimental neurological and muscular symptoms [13,14]. The first-line therapy for Gaucher is Glucocerebrosidase enzyme replacement therapy, that is a burdensome therapy due to the routine injections that the sufferers undertake. c-Rel Inhibitor Compound glucosylceramide synthase (GCS) is the GT that produces these glucocerebrosides applying UDP-Glucose as a donor and ceramides as acceptor substrates. An option to the intense enzyme replacement therapy, the identification of a little molecule inhibitor of GCS that could lower the glucosylceramide product in the brain and be administered orally, could possibly be a valuable treatment of Gaucher disease (Substrate reduction therapy) [15]. Because of the value of this class of enzymes, there is a have to have to create bioassays to study their activity and their regulation or determine chemical compounds that modulate their activity. Currently, measuring glycosyltransferase activity relies on standard methods, like the chromatographic separation of substrate and solution or the detection of a radiolabeled product. Although these assays have proved to be worthwhile when it comes to sensitivity and precision, they are cumbersome as they need washing methods and separation with the glycosylated solution for evaluation and aren’t simply configured for speedy screening [16]. Alternatively, various assay technologies not requiring the usage of radiochemicals had been developed inside the final two decades [17]. A few of them are fluorescence-based assays that detect the nucleoside diphosphate applying either fluorescent chemosensors [18,19] or fluorescent tracers combined with immunodetection [20]. These assays possess the advantage of getting universal for all GTs that release the detected nucleotide. Even so, specificity towards the nucleotide versus the nucleotide-sugar substrate can create larger background; hence, decreasing the sensitivity along with the accuracy from the assay. Furthermore, chemosensors’ availability and synthesis price could limit their widespread acceptance [17]. Other universal nucleotide detection assays relying on the enzyme-coupled generation of fluorescence or absorbance had been also developed for GT activity measurement [21,22]. The fluorescent GT assays rel