An investigation of the activity of recombinant rat skeletal muscle cytosolic sialidase.
Rat cytosolic sialidase is expressed at elevated levels in skeletal muscle and is believed to play a role in the myogenic differentiation of muscle cells. Here, we observed varying levels of enhancement of sialidase activity in the presence a range of divalent cations. In particular, a significant enhancement of activity was observed in the presence of Ca2+. Conversely, inhibition of the sialidase activity was found when the enzyme was incubated in the presence of Cu2+, EDTA, and a range of carbohydrate-based inhibitors. Finally, an investigation of the enzymatic hydrolysis of a synthetic substrate, 4-methylumbelliferyl N-acetyl-alpha-D-neuraminide, by 1H NMR spectroscopy revealed that the reaction catalysed by rat skeletal muscle cytosolic sialidase proceeds with overall retention of anomeric configuration. This result further supports the notion that all sialidases appear to be retaining enzymes.
Albouz-Abo
Samia
S
Department of Medicinal Chemistry, Monash University (Parkville Campus), 381 Royal Parade, Parkville, Vic. 3052, Australia.
Turton
Richard
R
Wilson
Jennifer C
JC
von Itzstein
Mark
M
D000818:Animals; D002241:Carbohydrates / Q000494:pharmacology; D002384:Catalysis; D002846:Chromatography, Affinity; D003600:Cytosol / Q000201:enzymology*; D006868:Hydrolysis; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D018482:Muscle, Skeletal / Q000201:enzymology*; D009439:Neuraminidase / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*
D002241:Carbohydrates; D011994:Recombinant Proteins; D009439:Neuraminidase