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Regulation ofN-acetylglucosaminyltransferase V by protein kinases

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Abstract

When 7721 human hepatocarcinoma cells were treated with 100nm phorbol-12-myristate-13-acetate (PMA), the activity ofN-acetylglucosaminyltransferase V(GnT-V) in the cells varied in accordance with the activity of membranous protein kinase C (PKC), but not with that of cytosolic PKC. Quercetin, a non-specific inhibitor of Ser/Thr protein kinase, andd-sphingosine and staurosporine, two specific inhibitors of PKC, blocked the activation of membranous PKC and GnT-V by PMA. Among the three inhibitors, quercetin was least effective. The inhibitory rates of quercetin and staurosporine toward membranous PKC and GnT V were proportional to the concentrations of the two inhibitors. The activities of GnT V and membranous protein kinase A (PKA) were also induced in parallel by dibutyryl cAMP (db-cAMP) and this induction was blocked by a specific PKA inhibitor. When cell free preparations of 7721 cells and human kidney were treated with alkaline phosphatase (ALP) to remove the phosphate groups, the GnT V activities were decreased. These results suggest that GnT V may be activated by membranous PKC or PKA, indirectly or directly, via phosphorylation of Ser/Thr residues.

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Abbreviations

UDP:

uridine diphospho-

GnT:

N-acetylglucosaminyltransferase

GlcNAc Gn:

N-acetylglucosamine

M:

mannose

PMA:

phorbol-12-myristate-13-acetate

PKC:

protein kinase C

PKA:

protein kinase A

cAMP:

adenosine 3′, 5′-cyclic monophosphate

db-cAMP:

dibutyryl cAMP

TPK:

tyrosine protein kinase

MES:

2-[N-morpholino]ethanesulfonic acid

DTT:

dithiothreitol

PMSF:

phenylmethylsulfonyl fluoride

EDTA:

ethylene diamine tetraacetic acid

EGTA:

glycol-bis-(α-aminoethyl) etherN,N,N′,N′-tetraacetic acid

PA:

2-aminopyridine

ALP:

alkaline phosphatase

C2C2 :

GlcNAcβ1-2 Manα1-6(GlcNAcβ1-2Manα1-3)ManβR

C2,4C2 :

GlcNAcβ1-2Manα1-6(GlcNAcβ1-4[GlcNAcβ1-2]Manα1-3)ManβR

C2C2,6 :

GlcNAcβ1-6[GlcNAcβ1-2]Manα1-6(GlcNAcβ1-2Manα1-3)ManβR

C2,4C2,6 :

GlcNAcβ1-6[GlcNAcβ1-2]Manα1-6(GlcNAcβ1-4[GlcNAcβ1-2]Manα1-3)ManβR where R=1-4GlcNAcβ1-4GlcNAcAsnX

Gn2M3Gn2-PA:

C2C2 where R=1-4GlcNAcβ1-4GlcNAc-PA

Gn3M3Gn2-PA:

C2C2,6 where R=1-4GlcNAcβ1-4GlcNAc-PA

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Authors and Affiliations

  1. Laboratory of Glycoconjugate Research, Department of Biochemistry, Shanghai Medical University, 200032, Shanghai, China

    Tong-Zhong Ju, Hui-Li Chen, Jian-Xin Gu & Hui Qin

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  1. Tong-Zhong Ju
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Ju, TZ., Chen, HL., Gu, JX. et al. Regulation ofN-acetylglucosaminyltransferase V by protein kinases. Glycoconjugate J 12, 767–772 (1995). https://doi.org/10.1007/BF00731237

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  • DOI: https://doi.org/10.1007/BF00731237

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