Neutrophils Exhibit Rapid Agonist-induced Increases in Protein-associated O-GlcNAc

A variety of cytoplasmic and nuclear proteins can be modified on serine and threonine residues by O-linked β-N-acetylglucosamine (O-GlcNAc), although the effects of this modification on protein and cellular functions are not completely defined. The sugar donor for the O-GlcNAc transferase that catalyzes this post-translational modification is UDP-N-acetylglucosamine (UDP-GlcNAc), a product of the hexosamine biosynthesis pathway (HBP). Here, the dynamics of the O-GlcNAc modification are examined in the physiological context of agonist-induced signal transduction using neutrophils. Formylated Met-Leu-Phe (fMLF) is shown to stimulate a rapid and transient increase in protein O-GlcNAcylation in both immunoblot and immunofluorescence imaging assays using O-GlcNAc-specific antibodies. In high performance liquid chromatography analyses of HBP metabolic activity, short term exposure to an exogenous substrate of the HBP, glucosamine (GlcNH2), leads to increased GlcNH2 6-phosphate and then UDP-GlcNAc levels. The GlcNH2 treatments also increase O-GlcNAcylation and augment the aforementioned fMLF-associated increase. In GlcNH2 on of O-GlcNAc in the of is to stimulate that a O-GlcNAcylation pathway that is by that O-GlcNAcylation an in rapid and signal to A variety of cytoplasmic and nuclear proteins can be modified on serine and threonine residues by O-linked β-N-acetylglucosamine (O-GlcNAc), although the effects of this modification on protein and cellular functions are not completely defined. The sugar donor for the O-GlcNAc transferase that catalyzes this post-translational modification is UDP-N-acetylglucosamine (UDP-GlcNAc), a product of the hexosamine biosynthesis pathway (HBP). Here, the dynamics of the O-GlcNAc modification are examined in the physiological context of agonist-induced signal transduction using neutrophils. Formylated Met-Leu-Phe (fMLF) is shown to stimulate a rapid and transient increase in protein O-GlcNAcylation in both immunoblot and immunofluorescence imaging assays using O-GlcNAc-specific antibodies. In high performance liquid chromatography analyses of HBP metabolic activity, short term exposure to an exogenous substrate of the HBP, glucosamine (GlcNH2), leads to increased GlcNH2 6-phosphate and then UDP-GlcNAc levels. The GlcNH2 treatments also increase O-GlcNAcylation and augment the aforementioned fMLF-associated increase. In GlcNH2 on of O-GlcNAc in the of is to stimulate that a O-GlcNAcylation pathway that is by that O-GlcNAcylation an in rapid and signal to The of in to residues O-linked GlcNH2 glucosamine HBP, hexosamine biosynthesis high performance liquid modification of O-linked GlcNH2 glucosamine HBP, hexosamine biosynthesis high performance liquid modification of is a post-translational modification that on cytoplasmic and nuclear proteins and is in the pathway modified in this nuclear and on this modification that and and to the that O-GlcNAc a to The and of for the O-GlcNAc transferase and of O-GlcNAc this the that to for O-GlcNAc and cellular and cellular in and O-GlcNAcylation to in the of and a in the of and of the to O-GlcNAc on the of hexosamine of the the hexosamine biosynthesis pathway to glucosamine 6-phosphate and UDP-N-acetylglucosamine (UDP-GlcNAc), the sugar donor for to be to in substrate a of substrate proteins of UDP-GlcNAc in in are to of O-GlcNAc that can be by HBP that of to HBP and UDP-GlcNAc and is not in the cellular is also and this in O-GlcNAc of O-GlcNAc an protein modification that O-GlcNAc of protein protein and The of in this and is O-GlcNAc can a signal transduction to the for the of to of the on the of O-GlcNAc on term high GlcNH2 the of that in the O-GlcNAcylation to this protein that O-GlcNAcylation can be a shown to in cellular O-GlcNAc short In that and rapid in In treatments the and the protein shown to in O-GlcNAc that O-GlcNAc are of the of O-GlcNAc a that is to of physiological in to in although is that are to an increase in and a increase in HBP a of on the O-GlcNAcylation to a and of a of in a variety of metabolic and a and in to this context for the signal transduction in cellular of the the to and protein of are to a of for to and the of to the of O-GlcNAc in cellular to using an that a HBP and O-GlcNAcylation In also of rapid and agonist-induced in that to both GlcNH2 and to and in protein the that protein O-GlcNAcylation is a of to cellular by and a of The for the and in by in for The then in and in The by the of in and for of by and to using a rapid for the in and The a of in for and then in O-GlcNAc-specific the The then a of in for in the The by an of for and in The for The in for and The for the for in and then a of in for O-GlcNAc-specific the the then for and then a of in for The and of to of nuclear of O-GlcNAc by using of and UDP-GlcNAc and for GlcNH2 and UDP-GlcNAc the and the of The for and of the The for and the for to of UDP-GlcNAc the on a the to The the of and for A and a to The for UDP-GlcNAc the of GlcNH2 of the of for by of The then a the to The the of and for A and a to by a of to The for GlcNH2 The and the by of of of protein and of GlcNH2 of UDP-GlcNAc and GlcNH2 to in using the for protein in modified and then in the of a The modified and in the of The in a for The then and by of by The and of GlcNH2 in the to by in a by in is a and a of signal transduction by a to signal protein of to increased O-GlcNAc by immunoblot analyses of the O-GlcNAc-specific A variety of proteins in the of to be for analyses that O-GlcNAcylation of and In the of by the O-GlcNAc-specific in immunoblot and assays not to a using in a an and of in O-GlcNAc increased O-GlcNAc of signal using by The a in this and high not The agonist-induced signal for by immunoblot rapid agonist-induced protein using the The of this for protein O-GlcNAc by to the for a and a and the O-GlcNAc to the the using the nuclear for of a that to the in the that are and and are the shown in the both the and in and The of a and and a in to a that the to the The cytoplasmic the increase in of the and the cytoplasmic although increased a variety of proteins are to be and a nuclear signal be is not this a of the a of proteins in a of a of the the this also to be a of the dynamics of agonist-induced in protein O-GlcNAcylation of and to for the that O-GlcNAc is a protein is an of the dynamics of agonist-induced protein using the and a nuclear cytoplasmic O-GlcNAcylation that for to levels. for to The of O-GlcNAc by of examined for in to by a HBP GlcNH2 to the for HBP an in O-GlcNAc the substrate to and O-GlcNAcylation that a and O-GlcNAcylation the effects of the HBP in GlcNH2 is an exogenous substrate for the HBP that is to O-GlcNAcylation a is by to GlcNH2 the the HBP product for examined GlcNH2 and UDP-GlcNAc in by GlcNH2 UDP-GlcNAc of protein of protein UDP-GlcNAc the sugar in and a and not that a of UDP-GlcNAc is for and treatments GlcNH2 to the of both GlcNH2 and GlcNH2 GlcNH2 and by that GlcNH2 the and by The increase in UDP-GlcNAc the in GlcNH2 be for HBP the increase in UDP-GlcNAc that for GlcNH2 the increase in UDP-GlcNAc that for GlcNH2 of of protein in cellular for GlcNH2 not In in GlcNH2 UDP-GlcNAc not exogenous GlcNH2 metabolic that are in the pathway to protein GlcNH2 in GlcNH2 for for on a using UDP-GlcNAc in GlcNH2 for for on a using O-GlcNAc the the in HBP in GlcNH2 for protein O-GlcNAc in by the O-GlcNAc-specific A of proteins O-GlcNAc a of in the to The to that by to A and The of in proteins modified by a of that not to increased O-GlcNAc in the The of for proteins assays using the that increase using the The of the for protein O-GlcNAc by to the O-GlcNAcylation by GlcNH2 for UDP-GlcNAc and O-GlcNAcylation analyses for GlcNH2 for and immunofluorescence using the The of this for protein O-GlcNAc by to the GlcNH2 for the increased UDP-GlcNAc and protein O-GlcNAc in the for a donor and and that in protein O-GlcNAcylation in UDP-GlcNAc and O-GlcNAcylation on the GlcNH2 and and UDP-GlcNAc and and an for O-GlcNAcylation of for although this not a of a for UDP-GlcNAc to to O-GlcNAc the shown in this for the and GlcNH2 an and of GlcNH2 for a increased signal The signal for by and immunoblot the increase in by to that for to A and The effects of GlcNH2 in the a increase in the The and in the and the also increased by GlcNH2 GlcNH2 in a of GlcNH2 and treatments for short GlcNH2 not increase O-GlcNAc by to to increased O-GlcNAc effects GlcNH2 also that the of O-GlcNAcylation to that for protein using the GlcNH2 for and The then for by for the The not and not shown that GlcNH2 can be to to this on fMLF-associated an of in O-GlcNAc a examined the of GlcNH2 on agonist-induced by the of and GlcNH2 of and on the by in assays a and for short GlcNH2 and and and for The GlcNH2 treatments not not not by for GlcNH2 for and and then not GlcNH2 In by of in assays for not a and GlcNH2 and of in the of and and in the of and for in assays for not and GlcNH2 for and to in and The not for of the of short GlcNH2 for on that GlcNH2 both and and in assays the that GlcNH2 of by both the of GlcNH2 and the of GlcNH2 The on the GlcNH2 and and and for GlcNH2 by of GlcNH2 of and O-GlcNAcylation is an modification that protein the physiological in rapid in the of O-GlcNAc are to be in O-GlcNAcylation are in the of a by and also to a in O-GlcNAc are by that O-GlcNAc to the metabolic of the and that O-GlcNAc and be to cellular In O-GlcNAcylation is also is in the of a variety of The of in to rapid in protein is to a for O-GlcNAc a and an that O-GlcNAc can be in a and a signal for signal for is of in O-GlcNAc this In modified a of a for this post-translational modification in signal and increased O-GlcNAc on a of The this modification in both immunofluorescence and in an increase in and a of proteins modified by O-GlcNAc in to immunofluorescence of the that the cytoplasmic the in the is not this is of O-GlcNAc is in a of the a variety of proteins are to be nuclear proteins and be that the of the the in the shown to be for O-GlcNAc by for the of the in the dynamics of In in in both immunoblot and immunofluorescence to for a of agonist-induced O-GlcNAcylation and the that short to that exogenous GlcNH2 be by the HBP in a term GlcNH2 treatments increased the of HBP GlcNH2 and then UDP-GlcNAc on the and of GlcNH2 treatments high GlcNH2 UDP-GlcNAc in O-GlcNAcylation GlcNH2 for although GlcNH2 increased O-GlcNAc the UDP-GlcNAc is a in UDP-GlcNAc and GlcNH2 in O-GlcNAcylation in UDP-GlcNAc of are the protein of shown to be to in UDP-GlcNAc In the of UDP-GlcNAc and not UDP-GlcNAc not be the of GlcNH2 the not that in cellular is of UDP-GlcNAc are to be also GlcNH2 and treatments and in in for high proteins in the short GlcNH2 by that GlcNH2 agonist-induced signal transduction by the of O-GlcNAcylation by the also GlcNH2 agonist-induced the UDP-GlcNAc and O-GlcNAc can be HBP to for O-GlcNAc The that HBP UDP-GlcNAc and also not that protein O-GlcNAc of agonist-induced the that fMLF-associated to and are The and of the that O-GlcNAc is a signal that in the context of physiological signal The of this post-translational the of proteins that are to be modified by and to in protein that a in in to cellular The of in to residues O-linked GlcNH2 glucosamine HBP, hexosamine biosynthesis high performance liquid modification of O-linked GlcNH2 glucosamine HBP, hexosamine biosynthesis high performance liquid modification of is a post-translational modification that on cytoplasmic and nuclear proteins and is in the pathway modified in this nuclear and on this modification that and and to the that O-GlcNAc a to The and of for the O-GlcNAc transferase and of O-GlcNAc this the that to for O-GlcNAc and cellular and cellular in and O-GlcNAcylation to in the of and a in the of and of the to O-GlcNAc on the of hexosamine of the the hexosamine biosynthesis pathway to glucosamine 6-phosphate and UDP-N-acetylglucosamine (UDP-GlcNAc), the sugar donor for to be to in substrate a of substrate proteins of UDP-GlcNAc in in are to of O-GlcNAc that can be by HBP that of to HBP and UDP-GlcNAc and is not in the cellular is also and this in O-GlcNAc of O-GlcNAc an protein modification that O-GlcNAc of protein protein and The of in this and is O-GlcNAc can a signal transduction to the for the of to The of the on the of O-GlcNAc on term high GlcNH2 the of that in the O-GlcNAcylation to this protein that O-GlcNAcylation can be a shown to in cellular O-GlcNAc short In that and rapid in In treatments the and the protein shown to in O-GlcNAc that O-GlcNAc are of the of O-GlcNAc a that is to of physiological in to in although is that are to an increase in and a increase in HBP a of on the O-GlcNAcylation to a and of a of in a variety of metabolic and a and in to this context for the signal transduction in cellular of the the to and protein of are to a of for to and the of to the of O-GlcNAc in cellular to using an that a HBP and O-GlcNAcylation In also of rapid and agonist-induced in that to both GlcNH2 and to and in protein the that protein O-GlcNAcylation is a of to cellular by and a of The for the and in by in for The then in and in The by the of in and for of by and to using a rapid for the in and The a of in for and then in O-GlcNAc-specific the The then a of in for in the The by an of for and in The for The in for and The for the for in and then a of in for O-GlcNAc-specific the the then for and then a of in for The and of to of nuclear of O-GlcNAc by using of and UDP-GlcNAc and for GlcNH2 and UDP-GlcNAc the and the of The for and of the The for and the for to of UDP-GlcNAc the on a the to The the of and for A and a to The for UDP-GlcNAc the of GlcNH2 of the of for by of The then a the to The the of and for A and a to by a of to The for GlcNH2 The and the by of of of protein and of GlcNH2 of UDP-GlcNAc and GlcNH2 to in using the for protein in modified and then in the of a The modified and in the of The in a for The then and by of by The and of GlcNH2 in the to by in a by by and a of The for the and in by in for The then in and in The by the of in and for of by and to using a rapid for the in and The a of in for and then in O-GlcNAc-specific the The then a of in for in the The by an of for and in The for The in for and The for the for in and then a of in for O-GlcNAc-specific the the then for and then a of in for The and of to of nuclear of O-GlcNAc by using of GlcNH2 and UDP-GlcNAc and for GlcNH2 and UDP-GlcNAc the and the of The for and of the The for and the for to of UDP-GlcNAc the on a the to The the of and for A and a to The for UDP-GlcNAc the of GlcNH2 of the of for by of The then a the to The the of and for A and a to by a of to The for GlcNH2 The and the by of of of protein and of GlcNH2 of UDP-GlcNAc and GlcNH2 to in using the for protein in modified and then in the of a The modified and in the of The in a for The then and by of by The and of GlcNH2 in the to by in a by in is a and a of signal transduction by a to signal protein of to increased O-GlcNAc by immunoblot analyses of the O-GlcNAc-specific A variety of proteins in the of to be for analyses that O-GlcNAcylation of and In the of by the O-GlcNAc-specific in immunoblot and assays not to a using in a an and of in O-GlcNAc increased O-GlcNAc of signal using by The a in this and high not The agonist-induced signal for by immunoblot both the and in and The of a and and a in to a that the to the The cytoplasmic the increase in of the and the cytoplasmic although increased a variety of proteins are to be and a nuclear signal be is not this a of the a of proteins in a of a of the the this also to be a of the dynamics of agonist-induced in protein O-GlcNAcylation of and to for the that O-GlcNAc is a protein is an of the dynamics of agonist-induced protein using the and a nuclear cytoplasmic O-GlcNAcylation that for to levels. for to The of O-GlcNAc by of examined for in to by a HBP GlcNH2 to the for HBP an in O-GlcNAc the substrate to and O-GlcNAcylation that a and O-GlcNAcylation the effects of the HBP in GlcNH2 is an exogenous substrate for the HBP that is to O-GlcNAcylation a is by to GlcNH2 the the HBP product for examined GlcNH2 and UDP-GlcNAc in by GlcNH2 UDP-GlcNAc of protein of protein UDP-GlcNAc the sugar in and a and not that a of UDP-GlcNAc is for and treatments GlcNH2 to the of both GlcNH2 and GlcNH2 GlcNH2 and by that GlcNH2 the and by The increase in UDP-GlcNAc the in GlcNH2 be for HBP the increase in UDP-GlcNAc that for GlcNH2 the increase in UDP-GlcNAc that for GlcNH2 of of protein in cellular for GlcNH2 not In in GlcNH2 UDP-GlcNAc not exogenous GlcNH2 metabolic that are in the pathway to protein GlcNH2 in GlcNH2 for for on a using UDP-GlcNAc in GlcNH2 for for on a using O-GlcNAc the the in HBP in GlcNH2 for protein O-GlcNAc in by the O-GlcNAc-specific A of proteins O-GlcNAc a of in the to The to that by to A and The of in proteins modified by a of that not to increased O-GlcNAc in the The of for proteins assays using the that increase using the The of the for protein O-GlcNAc by to the O-GlcNAcylation by GlcNH2 for UDP-GlcNAc and O-GlcNAcylation analyses for GlcNH2 for and immunofluorescence using the The of this for protein O-GlcNAc by to the GlcNH2 for the increased UDP-GlcNAc and protein O-GlcNAc in the for a donor and and that in protein O-GlcNAcylation in UDP-GlcNAc and O-GlcNAcylation on the GlcNH2 and and UDP-GlcNAc and and an for O-GlcNAcylation of for although this not a of a for UDP-GlcNAc to to O-GlcNAc the shown in this for the and GlcNH2 an and of GlcNH2 for a increased signal The signal for by and immunoblot the increase in by to that for to A and The effects of GlcNH2 in the a increase in the The and in the and the also increased by GlcNH2 GlcNH2 in a of GlcNH2 and treatments for short GlcNH2 not increase O-GlcNAc by to to increased O-GlcNAc effects GlcNH2 also that the of O-GlcNAcylation to that for protein using the GlcNH2 for and The then for by for the The not and not shown that GlcNH2 can be to to this on fMLF-associated an of in O-GlcNAc a examined the of GlcNH2 on agonist-induced by the of and GlcNH2 of and on the by in assays a and for short GlcNH2 and and and for The GlcNH2 treatments not not not by for GlcNH2 for and and then not GlcNH2 In by of in assays for not a and GlcNH2 and of in the of and and in the of and for in assays for not and GlcNH2 for and to in and The not for of the of short GlcNH2 for on that GlcNH2 both and and in assays the that GlcNH2 of by both the of GlcNH2 and the of GlcNH2 The on the GlcNH2 and and and for GlcNH2 by of GlcNH2 of and in is a and a of signal transduction by a to signal protein of to increased O-GlcNAc by immunoblot analyses of the O-GlcNAc-specific A variety of proteins in the of to be for analyses that O-GlcNAcylation of and In the of by the O-GlcNAc-specific in immunoblot and assays not to a using in a an and of in O-GlcNAc increased O-GlcNAc of signal using by The a in this and high not The agonist-induced signal for by immunoblot both the and in and The of a and and a in to a that the to the The cytoplasmic the increase in of the and the cytoplasmic although increased a variety of proteins are to be and a nuclear signal be is not this a of the a of proteins in a of a of the the this also to be a of the dynamics of agonist-induced in protein O-GlcNAcylation of and to for the that O-GlcNAc is a protein a HBP GlcNH2 to the for HBP an in O-GlcNAc the substrate to and O-GlcNAcylation that a and O-GlcNAcylation the effects of the HBP in GlcNH2 is an exogenous substrate for the HBP that is to O-GlcNAcylation a is by to GlcNH2 the the HBP product for examined GlcNH2 and UDP-GlcNAc in by GlcNH2 UDP-GlcNAc of protein of protein UDP-GlcNAc the sugar in and a and not that a of UDP-GlcNAc is for and treatments GlcNH2 to the of both GlcNH2 and GlcNH2 GlcNH2 and by that GlcNH2 the and by The increase in UDP-GlcNAc the in GlcNH2 be for HBP the increase in UDP-GlcNAc that for GlcNH2 the increase in UDP-GlcNAc that for GlcNH2 of of protein in cellular for GlcNH2 not In in GlcNH2 UDP-GlcNAc not GlcNH2 O-GlcNAc the the in HBP in GlcNH2 for protein O-GlcNAc in by the O-GlcNAc-specific A of proteins O-GlcNAc a of in the to The to that by to A and The of in proteins modified by a of that not to increased O-GlcNAc in the The of for proteins assays using the increased UDP-GlcNAc and protein O-GlcNAc in the for a donor and and that in protein O-GlcNAcylation in UDP-GlcNAc and O-GlcNAcylation on the GlcNH2 and and UDP-GlcNAc and and an for O-GlcNAcylation of for although this not a of a for UDP-GlcNAc to to O-GlcNAc the shown in this for the and GlcNH2 an and of GlcNH2 for a increased signal The signal for by and immunoblot the increase in by to that for to A and The effects of GlcNH2 in the a increase in the The and in the and the also increased by GlcNH2 GlcNH2 GlcNH2 in a of GlcNH2 and treatments for short GlcNH2 not increase O-GlcNAc by to to increased O-GlcNAc effects GlcNH2 also that the of O-GlcNAcylation to that for GlcNH2 shown that GlcNH2 can be to to this on fMLF-associated an of in O-GlcNAc a examined the of GlcNH2 on agonist-induced by the of and GlcNH2 of and on the by in assays a and for short GlcNH2 and and and for The GlcNH2 treatments not not of the of short GlcNH2 for on that GlcNH2 both and and in assays the that GlcNH2 of by both the of GlcNH2 and the of GlcNH2 The on the GlcNH2 and and and for GlcNH2 by of GlcNH2 of and O-GlcNAcylation is an modification that protein the physiological in rapid in the of O-GlcNAc are to be in O-GlcNAcylation are in the of a by and also to a in O-GlcNAc are by that O-GlcNAc to the metabolic of the and that O-GlcNAc and be to cellular In O-GlcNAcylation is also is in the of a variety of The of in to rapid in protein is to a for O-GlcNAc a and an that O-GlcNAc can be in a and a signal for signal for is of in O-GlcNAc this In modified a of a for this post-translational modification in signal and increased O-GlcNAc on a of The this modification in both immunofluorescence and in an increase in and a of proteins modified by O-GlcNAc in to immunofluorescence of the that the cytoplasmic the in the is not this is of O-GlcNAc is in a of the a variety of proteins are to be nuclear proteins and be that the of the the in the shown to be for O-GlcNAc by for the of the in the dynamics of In in in both immunoblot and immunofluorescence to for a of agonist-induced O-GlcNAcylation and the that short to that exogenous GlcNH2 be by the HBP in a term GlcNH2 treatments increased the of HBP GlcNH2 and then UDP-GlcNAc on the and of GlcNH2 treatments high GlcNH2 UDP-GlcNAc in O-GlcNAcylation GlcNH2 for although GlcNH2 increased O-GlcNAc the UDP-GlcNAc is a in UDP-GlcNAc and GlcNH2 in O-GlcNAcylation in UDP-GlcNAc of are the protein of shown to be to in UDP-GlcNAc In the of UDP-GlcNAc and not UDP-GlcNAc not be the of GlcNH2 the not that in cellular is of UDP-GlcNAc are to be also GlcNH2 and treatments and in in for high proteins in the short GlcNH2 by that GlcNH2 agonist-induced signal transduction by the of O-GlcNAcylation by the also GlcNH2 agonist-induced the UDP-GlcNAc and O-GlcNAc can be HBP to for O-GlcNAc The that HBP UDP-GlcNAc and also not that protein O-GlcNAc of agonist-induced the that fMLF-associated to and are The and of the that O-GlcNAc is a signal that in the context of physiological signal The of this post-translational the of proteins that are to be modified by and to in protein that a in in to cellular O-GlcNAcylation is an modification that protein the physiological in rapid in the of O-GlcNAc are to be in O-GlcNAcylation are in the of a by and also to a in O-GlcNAc are by that O-GlcNAc to the metabolic of the and that O-GlcNAc and be to cellular In O-GlcNAcylation is also is in the of a variety of The of in to rapid in protein is to a for O-GlcNAc a and an that O-GlcNAc can be in a and a signal for The signal for is of in O-GlcNAc this In modified a of a for this post-translational modification in signal and increased O-GlcNAc on a of The this modification in both immunofluorescence and in an increase in and a of proteins modified by O-GlcNAc in to immunofluorescence of the that the cytoplasmic the in the is not this is of O-GlcNAc is in a of the a variety of proteins are to be nuclear proteins and be that the of the the in the shown to be for O-GlcNAc by for the of the in the dynamics of In in in both immunoblot and immunofluorescence to for a of agonist-induced O-GlcNAcylation and the that short to that exogenous GlcNH2 be by the HBP in a term GlcNH2 treatments increased the of HBP GlcNH2 and then UDP-GlcNAc on the and of GlcNH2 treatments high GlcNH2 UDP-GlcNAc in O-GlcNAcylation GlcNH2 for although GlcNH2 increased O-GlcNAc the UDP-GlcNAc is a in UDP-GlcNAc and GlcNH2 in O-GlcNAcylation in UDP-GlcNAc of are the protein of shown to be to in UDP-GlcNAc In the of UDP-GlcNAc and not UDP-GlcNAc not be the of GlcNH2 the not that in cellular is of UDP-GlcNAc are to be also GlcNH2 and treatments and in in for high proteins in the short GlcNH2 by that GlcNH2 agonist-induced signal transduction by the of O-GlcNAcylation by the also GlcNH2 agonist-induced the UDP-GlcNAc and O-GlcNAc can be HBP to for O-GlcNAc The that HBP UDP-GlcNAc and also not that protein O-GlcNAc of agonist-induced the that fMLF-associated to and are The and of In the that O-GlcNAc is a signal that in the context of physiological signal The of this post-translational the of proteins that are to be modified by and to in protein that a in in to cellular for and for and the of and for the of the and and the for to immunofluorescence