TY - JOUR
T1 - Cellular metabolism of unnatural sialic acid precursors
AU - Pham, Nam D.
AU - Fermaintt, Charles S.
AU - Rodriguez, Andrea C.
AU - McCombs, Janet E.
AU - Nischan, Nicole
AU - Kohler, Jennifer J.
N1 - Publisher Copyright:
© 2015 Springer Science+Business Media New York.
PY - 2015/10/1
Y1 - 2015/10/1
N2 - Carbohydrates, in addition to their metabolic functions, serve important roles as receptors, ligands, and structural molecules for diverse biological processes. Insight into carbohydrate biology and mechanisms has been aided by metabolic oligosaccharide engineering (MOE). In MOE, unnatural carbohydrate analogs with novel functional groups are incorporated into cellular glycoconjugates and used to probe biological systems. While MOE has expanded knowledge of carbohydrate biology, limited metabolism of unnatural carbohydrate analogs restricts its use. Here we assess metabolism of SiaDAz, a diazirine-modified analog of sialic acid, and its cell-permeable precursor, Ac4ManNDAz. We show that the efficiency of Ac4ManNDAz and SiaDAz metabolism depends on cell type. Our results indicate that different cell lines can have different metabolic roadblocks in the synthesis of cell surface SiaDAz. These findings point to roles for promiscuous intracellular esterases, kinases, and phosphatases during unnatural sugar metabolism and provide guidance for ways to improve MOE.
AB - Carbohydrates, in addition to their metabolic functions, serve important roles as receptors, ligands, and structural molecules for diverse biological processes. Insight into carbohydrate biology and mechanisms has been aided by metabolic oligosaccharide engineering (MOE). In MOE, unnatural carbohydrate analogs with novel functional groups are incorporated into cellular glycoconjugates and used to probe biological systems. While MOE has expanded knowledge of carbohydrate biology, limited metabolism of unnatural carbohydrate analogs restricts its use. Here we assess metabolism of SiaDAz, a diazirine-modified analog of sialic acid, and its cell-permeable precursor, Ac4ManNDAz. We show that the efficiency of Ac4ManNDAz and SiaDAz metabolism depends on cell type. Our results indicate that different cell lines can have different metabolic roadblocks in the synthesis of cell surface SiaDAz. These findings point to roles for promiscuous intracellular esterases, kinases, and phosphatases during unnatural sugar metabolism and provide guidance for ways to improve MOE.
KW - Diazirine
KW - Esterase
KW - Metabolic engineering
KW - Photocrosslinking
KW - Sialic acid
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U2 - 10.1007/s10719-015-9593-7
DO - 10.1007/s10719-015-9593-7
M3 - Article
C2 - 25957566
AN - SCOPUS:84944281146
SN - 0282-0080
VL - 32
SP - 515
EP - 529
JO - Glycoconjugate Journal
JF - Glycoconjugate Journal
IS - 7
ER -