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2.4.1.144: beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase

This is an abbreviated version!
For detailed information about beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase, go to the full flat file.

Word Map on EC 2.4.1.144

Reaction

UDP-N-acetyl-alpha-D-glucosamine
+
beta-D-GlcNAc-(1->2)-alpha-D-Man-(1->3)-[beta-D-GlcNAc-(1->2)-alpha-D-Man-(1->6)]-beta-D-Man-(1->4)-beta-D-GlcNAc-(1->4)-beta-D-GlcNAc-N-Asn-[protein]
=
UDP
 +
beta-D-GlcNAc-(1->2)-alpha-D-Man-(1->3)-[beta-D-GlcNAc-(1->2)-alpha-D-Man-(1->6)]-[beta-D-GlcNAc-(1->4)]-beta-D-Man-(1->4)-beta-D-GlcNAc-(1->4)-beta-D-GlcNAc-N-Asn-[protein]

Synonyms

(GnT)-III, acetylglucosaminyltransferase, uridine diphosphoacetylglucosamine-glycopeptide beta4-, III, beta(1,4)-N-acetylglucosaminyltransferase III, beta-1,4-mannosyl-glycoprotein beta-1,4-N-acetylglucosaminyltransferase, beta-1,4-N-acetylglucosaminyltransferase III, beta-D-mannoside beta-1,4-N-acetylglucosaminyltransferase, beta1,4-N-acetylglucosaminyltransferase III, EC 2.4.1.51, GlcNAc-transferase-III, GlcNAcTase-III, GnT-III, GnTIII, Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase III, MGAT3, N-acetyl-glucosaminyltransferase III, N-acetylglucosaminyltransferase III, N-acetylglucosaminyltransferase-III, N-glycosyl-oligosaccharide-glycoprotein N-acetylglucosaminyltransferase III, uridine diphosphate (UDP)-N-acetylglucosamin/beta-D-mannoside beta-1,4-N-acetylglucosaminyltransferase III, uridine diphosphoacetylglucosamine-glycopeptide beta4-acetylglucosaminyltransferase III

ECTree

     2 Transferases
         2.4 Glycosyltransferases
             2.4.1 Hexosyltransferases
                2.4.1.144 beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase

Cloned

Cloned on EC 2.4.1.144 - beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase

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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
amplified for cloning into pENTR-D-Topo vector, cloned gene inserted into the virus expression vector, pBABE-puro. GnT-III and GnT-V constructs transfected into Phoenix-Ampho cells
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cloning from genetic library, DNA sequence determination, functional overexpression in COS-1 or HeLa-cells, transient transfection
cloning of cDNA by RT-PCR
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construction of genes for expression of native rat GnTIII, human ManII, Arabidopsis thaliana ManII and for the expression of the catalytic domains of rat GnTIII and human ManII fused to the N-terminal region of Arabidopsis thaliana ManII. Binary plasmids for high-level expression of rat GnTIII (pAX67), chimeric rat GnTIIIA.th. (replaced native localization domain with the cytoplasmic tail, transmembrane, and stem region of Arabidopsis thaliana mannosidase II) (pAX70), co-expression of rat GnTIII together with human ManII (pAX73) or Arabidopsis thaliana ManII (pAX100), co-expression of GnTIIIA.th. together with ManIIA.th. (pAX74), and co-expression of GnTIIIA.th. and Arabidopsis thaliana ManII (pAX101). Expression of the transgenes is driven by a chimeric promoter assembled from regulatory elements from mannopine and octopine synthase genes. Expression cassettes comprising the CaMV 35S promoter, GnTIII or ManII and the nos pA, being pSK103 (ManII), pSK124 (ManII-Arabidopsis thaliana), pAX63 (GnTIII-Arabidopsis thaliana) and pAX68 (GnTIII) serve as starting point for further expression constructs. The cassette comprising the CaMV 35 promoter, plant relocalized GnTIII and polyadenylation signal is excised from pAX68 using SbfI, EcoRI and inserted into pAX36 generating pAX90. The construct comprising the rat GnTIII obtained by replacing the XbaI, AflII fragment from pAX90 with the fragment isolated from pAX63, generating pAX91. Mutated fragment from pCLF40 inserted into pAX69 using AvaI, StuI, generating pAX104. The binary expression plasmids for inactive GnTIII and GnTIIIA.th. under the control of the UAS123mas promoter generated by replacing the StuI, EcoRI fragment from pAX104 with that in pAX67 (generating pAX105) and pAX70 (generating pAX106), respectively. The mutated fragment from pCLF40 inserted into pAX91 using Eam1105I, StuI, generating the expression plasmid for inactive GnTIII under the control of the CaMV 35S promoter (pAX108). The exchange of the fragment containing the Arabidopsis thaliana Golgi localization domain from pAX90 in pAX108 using SbfI, StuI yields an expression plasmid for inactive GnTIIIA.th. under the control of the CaMV 35S promoter. Plasmids transferred into Agrobacterium tumefaciens LBA4404 for transformation into tobacco BY-2 cells
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construction of transgenic mice, disruption of certain functions of apolipoprotein B leading to generation of a aftty liver
enzyme overexpression in HeLa-S3, CHO-K1, Pro-5, and MDA-MB-231 cells using vector CSIV-TRE-RfA-CMV-KT, cloning in HEK-293T cells
expressed in Spodoptera frugiperda Sf21 cells
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expressed in transgenic mice
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expression in erythroleukemia cell line K562, leading to increased resistance to lysis by natural killer cells and enhanced spleen colonization ability
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expression of a catalytically inactive D232A mutant in human hepatoblastoma cell line Huh6, leading to suppression of the endogenous enzyme activity, but not to a significant decrease in the expression level of the endogenous enzyme
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expression of mutant aglycosyl-enzyme, lacking the first 23 amino acid residues, in Escherichia coli
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expression of soluble His-tagged protein in Spodoptera frugiperda Sf 21 insect cells via baculovirus infection, secretion into the medium
expression of wild-type and mutants in COS-1 cells
gene MGAT3, quantitative real-time PCR enzyme expression analysis
gene MGAT3, real-time RT-PCR enzyme expression analysis, establishing of human melanoma cell line WM-266-4 with induced stable overexpression of GnT-III, the cells show increased amounts of bisected and beta1-6 branched N-glycans on melanoma cell adhesion molecule MCAM/MUC18, but do not display significant differences in viability and capabilities to migrate through an endothelial layer, coexpression with MGAT5 gene, EC 2.4.1.155
gene MGAT3, stable overexpression of the enzyme in WM-266-4-GnT-III human metastatic melanoma cells
GnT-III overexpressed in MKN45 cells
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GnT-III stable expressing cell lines of Neuro-2a mouse neuroblastoma cells and B16 mouse melanoma cells are established. Overexpression of GnT-III up-regulates adenylyl cyclases activity in Neuro-2a mouse neuroblastoma cells and B16 mouse melanoma cells
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GnT-III transfection into highly pigmented, highly metastatic macrophage-melanoma fusion hybrids results in the suppression of both melanogenesis and motility. The effects of GnT-III appear to occur through inhibition of GnT-V action
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heterozygous alpha 1,3-galactosyltransferase gene knockout pigs produced with transgenic pig fetal cells expressing both human decay-accelerating factor and N-acetylglucosaminyltransferase III
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highly metastatic melanoma B16 cells transfected with GnT-III and then injected into syngeneic mice via the tail vein
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introduction of human N-acetylglucosaminyltransferase (GnT)-III gene into Nicotiana tabacum leads to highly efficient synthesis of bisected N-glycans
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introduction of human N-acetylglucosaminyltransferase gene into tobacco plants leads to highly efficient synthesis of bisected N-glycans. The majority of N-glycans of an antibody produced in a plant expressing GnT-III is also bisected. This might improve the efficacy of therapeutic antibodies produced in this type of transgenic plant
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overexpression in HeLa cells, leading to suppression of H2O2-induced activation of PKCdelta-JNK1 signalling pathway resulting in inhibition of apoptosis
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overexpression in murine lung melanoma cell line B16-hm, leading to suppression of the formation of beta-1,6-tri- and tetraantennary N-linked oligosaccharides by inhibiting the responsible enzymes, e.g. N-acetylglucosamine transferase V, EC 2.4.1.155, decrease of the metastatic potential of B16-hm melanoma in vivo, changed phenotype
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overexpression in rat pheochromocytoma cell line PC-12, glycoproteins contain elevated levels of bisecting GlcNAc, abolished cell differentiation
rat GnTIII is expressed either with its native localization domain (GnTIII) or with the cytoplasmic tail, transmembrane domain and stem region (CTS) of Arabidopsis thaliana mannosidase II. N-Glycans of plants expressing rat GnTIII contain three major glycan structures of complex bisected, complex, or hybrid bisected type, accounting for 70%-85% of the total N-glycans. Expression of rat beta(1,4)-N-acetylglucosaminyltransferase III in Nicotiana tabacum remodels the plant-specific N-glycosylation
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Sf21 cells infected with recombinant baculovirus encoding GnT-III
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