Hyaluronan (HA) is the main component of the extracellular matrix (ECM). Depending on its chain size, it is generally accepted to exert diverse effects. High molecular weight HA is anti-angiogenic, immunosuppressive and anti-inflammatory, while lower fragments are angiogenic and inflammatory. Human hyaluronidase Hyal-1 (Hyal-1) is one of the main enzymes in the metabolism of HA. This makes Hyal-1 an interesting target. Not only for functional and mechanistic studies, but also for drug development. In this work, Hyal-1 was expressed on the surface of E. coli, by applying Autodisplay, to overcome formation of inactive "inclusion bodies". With the cells displaying Hyal-1 an activity assay was performed OPEN ACCESSMolecules 2015, 20 15450 using "stains-all" dye. Subsequently, the inhibitory effects of four saponins and 14 plant extracts on the activity of surface displayed Hyal-1 were evaluated. The determined IC50 values were 177 µM for glycyrrhizic acid, 108 µM for gypsophila saponin 2, 371 µM for SA1657 and 296 µM for SA1641. Malvae sylvestris flos, Equiseti herba and Ononidis radix extracts showed IC50 values between 1.4 and 1.7 mg/mL. In summary, Autodisplay enabled the expression of functional human target protein Hyal-1 in E. coli and facilitated an accelerated testing of potential inhibitors.
Abstract:The negatively charged polysaccharide Hyaluronic acid (HA) has diverse physiological and pathophysiological functions depending on its chain size. Space filling, anti inflammatory and antiangiogenic effects are triggered by high molecular weight HA (HMW HA) (>20 kDa). Hydrolyzation of HMW HA by Hyal-1 results in low molecular weight HA (LMW HA) (<20 kDa) which leads to inflammatory and angiogenic effects.[1] For this reason Hyal-1 is an interesting target for drug discovery. The surface display of active Hyal-1 on Escherichia coli, via Autodisplay, enables the screening for potential inhibitors in a whole cell system. Based on this technique we determined the inhibitory effect of different natural substances on human Hyal-1. The IC 50 values of the plant extracts Malvae sylvestris flos, Equiseti herba and Ononidis radix were determined to be between 1.4 and 1.7 mg/mL. Furthermore, the IC 50 values of four triterpenoid saponines were determined. The obtained IC 50 value for glycyrrhizic acid, a known Hyal-1 inhibitor, was 177 µM. The IC 50 values for the newly identified inhibitors gypsophila saponin 2, SA1641, and SA1657 were 108 µM, 296 µM and 371 µM, respectively.[2] For the synthesis of new small molecule inhibitors targeting human Hyal-1 these extracts and natural compounds could be used as a starting point.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.