Biochemical engineering of the N-acyl side chain of sialic acid leads to increased calcium influx from intracellular compartments and promotes differentiation of HL60 cells.
Sialylation of glycoconjugates is essential for mammalian cells. Sialic acid is synthesized in the cytosol from N-acetylmannosamine by several consecutive steps. Using N-propanoylmannosamine, a novel precursor of sialic acid, we are able to incorporate unnatural sialic acids with a prolonged N-acyl side chain (e.g., N-propanoylneuraminic acid) into glycoconjugates taking advance of the cellular sialylation machinery. Here, we report that unnatural sialylation of HL60-cells leads to an increased release of intracellular calcium after application of thapsigargin, an inhibitor of SERCA Ca2+-ATPases. Furthermore, this increased intracellular calcium concentration leads to an increased adhesion to fibronectin. Finally, we observed an increase of the lectin galectin-3, a marker of monocytic differentiation of HL60-cells.
Horstkorte
Rüdiger
R
Institut für Biochemie und Molekularbiologie, Charité-Universitätsmedizin Berlin, Campus Benjamin Franklin, Arnimallee 22, 14195 Berlin-Dahlem, Germany. ruediger.horstkorte@charite.de
Rau
Kirstin
K
Laabs
Stephan
S
Danker
Kerstin
K
Reutter
Werner
W
D000215:Acylation; D002118:Calcium / Q000378:metabolism*; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors; D002454:Cell Differentiation / Q000502:physiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006001:Glycoconjugates / Q000378:metabolism; D018922:HL-60 Cells; D006801:Humans; D019158:N-Acetylneuraminic Acid / Q000378:metabolism*; D019284:Thapsigargin / Q000494:pharmacology
D004791:Enzyme Inhibitors; D006001:Glycoconjugates; D019284:Thapsigargin; D000252:Calcium-Transporting ATPases; D019158:N-Acetylneuraminic Acid; D002118:Calcium