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Enzyme Activity Measurement for Hyaluronan Synthase Using Chromatographic Assays

With years of discovering and developing, Creative Enzymes has emerged as a worldwide leader in enzyme activity assays. Creative Enzymes continues growing with the same entrepreneurial and innovative spirit bringing nature to science. As an enzyme manufacturer, Creative Enzymes always meets the requirements of our customers supporting the next development stage and therefore has been the best choice with satisfying services. As one of the highlighted services, we offer the accurate activity measurement for hyaluronan synthase.

Hyaluronan synthases (EC; HASs) are the membrane-bound glycosyltransferases responsible for the biosynthesis of hyaluronan (HA). They are dual-action enzymes that bind both UDP-α-D-glucuronic acid and UDP-α-N-acetyl-D-glucosamine in two different active sites that alternately catalyze the formation of two different glycosidic bonds in the presence of either Mg2+ or Mn2+. In mammals, HA is produced by three homologous HAS enzymes: HAS1, HAS2, and HAS3. These enzymes are highly specific for UDP-GlcNAc and UDP-GlcA, and similar enzymes have been found in a variety of organisms.

HA is a glycosaminoglycan polymer composed of repeated N-acetylglucosamine β(1,4) glucuronic acid β(1,3) disaccharide units. It traps a large amount of water, giving tissues high viscosity and elasticity. HA plays a pivotal role during fertilization, embryogenesis, development, and differentiation. Moreover, HA also participates in a wide range of cellular functions and behaviors, including cell migration, phagocytosis, and proteoglycan assembly. Furthermore, HA serves as a critical role in tissue remodeling, normal tissue homeostasis, and diseases, such as osteoarticular pathology, inflammatory disorders, and vascular diseases. In addition, HA also can be used as a drug delivery vehicle, a cosmetic ingredient, and as an analgesic. Considering the multiple functions of HA, HASs are the important enzymes in all of biological activity because the enzymes directly act on the biosynthesis of HA.

Although HASs have been characterized enzymologically, there are few reported studies about their structure and mechanism. One of the reasons for the limited amount of mechanistic information is the lack of a convenient activity assay. Here, Creative Enzymes is fully competent to provide the precise activity assay for HASs. HAS activity is monitored using UDP-[14C]GlcA by chromatography. The incorporation of UDP-[14C]GlcA into high molecular mass HA was measured by descending paper chromatography.

Our research center stands out with the most advanced equipment, a long research history, and an extraordinary expert team, which assure the high reproducibility and accuracy of the activity measurement. We deliver the products in the shortest span of time from the date of order placement. In the future, Creative Enzymes will continue to support your business with our specialized services.

Figure: The  crystal structure of human hyaluronan synthase 2.
Figure: The crystal structure of human hyaluronan synthase 2.
UniProt: Q92819

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CAS No.39346-43-5
SourceE. coli
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