Despite the fundamental roles of sialyl- and fucosyltransferases in mammalian physiology, there are few pharmacological tools to manipulate their function in a cellular setting. Although fluorinated analogs of the donor substrates are well-established transition state inhibitors of these enzymes, they are not membrane permeable. By exploiting promiscuous monosaccharide salvage pathways, we show that fluorinated analogs of sialic acid and fucose can be taken up and metabolized to the desired donor substrate–based inhibitors inside the cell. Because of the existence of metabolic feedback loops, they also act to prevent the de novo synthesis of the natural substrates, resulting in a global, family-wide shutdown of sialyl- and/or fucosyltransferases and remodeling of cell-surface glycans. As an example of the functional consequences, the inhibitors substantially reduce expression of the sialylated and fucosylated ligand sialyl Lewis X on myeloid cells, resulting in loss of selectin binding and impaired leukocyte rolling.
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This work was supported by US National Institutes of Health grants to J.C.P. (R01AI050143 and P01HL107151), C.D.R. (T32AI007606), K.L. (R01HL111969), C.T.L. (T32AI060536) and the Complex Carbohydrate Research Center (1 P41 RR018502-01) as well as by funding from the Biotechnology and Biological Sciences Research Council to A.D. and S.M.H. (BBF0083091).
The authors declare no competing financial interests.
Supplementary Methods and Supplementary Results (PDF 2406 kb)
Rolling Velocity Measurement of control (DMSO) treated HL-60 cells on E-Selectin at 3 dynes/cm2 (MOV 2837 kb)
Rolling Velocity Measurement of 2F-Fuc (2) treated HL-60 cells on ESelectin at 3 dynes/cm2 (MOV 3188 kb)
Rolling Velocity Measurement of 3Fax-Neu5Ac (8) treated HL-60 cells on E-Selectin at 3 dynes/cm2 (MOV 3169 kb)
Rolling Velocity Measurement of control (DMSO) treated HL-60 cells on P-Selectin at 3 dynes/cm2 (MOV 3596 kb)
Rolling Velocity Measurement of 2F-Fuc (2) treated HL-60 cells on PSelectin at 3 dynes/cm2 (MOV 3855 kb)
Rolling Velocity Measurement of 3Fax-Neu5Ac (8) treated HL-60 cells on P-Selectin at 3 dynes/cm2 (MOV 3516 kb)
Tethering Analysis of Control (DMSO) treated HL-60 cells on E-Selectin (MOV 2935 kb)
Tethering Analysis of 2F-Fuc (2) treated HL-60 cells on E-Selectin (MOV 2803 kb)
Tethering Analysis of 3Fax-Neu5Ac (8) treated HL-60 cells on E-Selectin (MOV 3301 kb)
Tethering Analysis of Control (DMSO) treated HL-60 cells on PSelectin (MOV 3605 kb)
Tethering Analysis of 2F-Fuc (2) treated HL-60 cells on P-Selectin (MOV 3842 kb)
Tethering Analysis of 3Fax-Neu5Ac (8) treated HL-60 cells on PSelectin (MOV 3778 kb)
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Rillahan, C., Antonopoulos, A., Lefort, C. et al. Global metabolic inhibitors of sialyl- and fucosyltransferases remodel the glycome. Nat Chem Biol 8, 661–668 (2012). https://doi.org/10.1038/nchembio.999
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