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Polyethylene Glycol Crowding Effect on Hyaluronidase Activity Monitored by Capillary Electrophoresis by Nasreddine, Rouba; Orlic, Lucija; Banni, Ghassan Al Hamoui Dit; Fayad, Syntia; Marchal, Axel; Piazza, Francesco; Lopin-Bon, Chrystel; Hamacek, Josef; Nehmé, Reine is a Chemistry article available to read on EtoBox.
What is Polyethylene Glycol Crowding Effect on Hyaluronidase Activity Monitored by Capillary Electrophoresis about?
To mimic the activity of hyaluronidase in natural environment, the hydrolysis of hyaluronic acid (HA) by hyaluronidase was investigated for the first time in the presence of crowding agents using capillary electrophoresis (CE) as a simple and reliable technique for conducting enzymatic assay. Polyethylene glycol (PEG) 6000 was selected as a model crowder and the hyaluronic acid degradation catalyzed by bovine testes hyaluronidase (BTH) was carried out at different PEG concentrations (0%, 10%, and 17%). After optimization of the CE analytical method and enzymatic assay, the degradation products were monitored at different HA concentrations. At 10% of PEG and 0.3 mg mL of HA, the activity of the enzyme was significantly reduced showing inconvenient interactions of PEG with the hyaluronidase blocking the release of hydrolysis products. A similar reduction of hyaluronidase activity was observed at 1 mg mL of HA due to the presumable formation of the BTH-substrate complex. The experimental curves obtained by CE also evidence that the overall kinetics are governed by the hydrolysis of hexasaccharide intermediates. Finally, the effect of PEG on hyaluronidase activity was evaluated in the
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- Author
- Nasreddine, Rouba; Orlic, Lucija; Banni, Ghassan Al Hamoui Dit; Fayad, Syntia; Marchal, Axel; Piazza, Francesco; Lopin-Bon, Chrystel; Hamacek, Josef; Nehmé, Reine
- Publisher
- Springer; Springer-Verlag; Springer Verlag; Springer Science and Business Media LLC; Society for Mining, Metallurgy and Exploration Inc.; Springer Nature (ISSN 1618-2650)
- Published
- 2020
- Language
- EN
- Field
- Chemistry (Physical Sciences)