This work investigated the substrate specificity of hNEU enzymes for a glycoprotein substrate (bovine submaxillary mucin) containing 9-O-acetylated and Neu5Gc residues. Using this model substrate, we observe a general trend for hNEU tolerance of Neu5Ac>Neu5Gc>>>Neu5,9Ac 2 , consistent with our previous results with glycolipid substrates. These results expand our understanding of hNEU enzyme specificity and suggest that naturally occurring modifications of sialic acids can play a role in regulating hNEU activity. File list (2) download file view on ChemRxiv hunter.text.rev.pdf (486.29 KiB) download file view on ChemRxiv hunter.si.pdf (288.03 KiB) Human neuraminidases have reduced activity towards modified sialic acids on glycoproteins
Gangliosides are important signaling molecules in the cell membrane and are processed by several enzymes. Deficiencies in these enzymes can cause human lysosomal storage diseases.Building an understanding of the pathwaysof glycosphingolipid catabolism requires methods for the analysis of these enzymatic activities AGM3-derived FRET probe was synthesized chemoenzymatically for the detection and quantitation of arange of ganglioside-degrading enzymes,both in cell lysates and in living cells.This is the first substrate that enables the ratiometric fluorogenic assayo fs phingolipid ceramide Ndeacylase and endoglycoceramidase and can detect and localizen euraminidase activity in living cells.I ti st herefore av aluable tool for building ab etter understanding of membrane-confined enzymology.I ta lso enables the robust and reliable assay of ganglioside-degrading enzymes in am icrotiter plate,t hus opening the door to screening for novel or engineered biocatalysts or for new inhibitors.
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.