| Organism Information |
| Organism Name | Actinobacillus pleuropneumoniae serotype 7 (strain AP76) |
| Clinical Implication | Pathogenic |
| Domain | Bacteria |
| Phylum | Proteobacteria |
| Classification | Family: Pasteurellaceae
Order: Pasteurellales
Class: Gammaproteobacteria
Phylum: Proteobacteria |
| Taxonomic ID (NCBI) | 537457 |
| Genome Information |
| Gene Bank | CP001091.1 |
| EMBL | CP001091.1 |
| Gene Information |
| Gene Name | APP7_1697 |
| Protein information |
| Protein Name | ApNGTÂ |
| UniProtKB/ SwissProt ID | B3H2N2 |
| NCBI Ref Seq | WP_012478567.1 |
| UniProtKB Sequence | >sp|B3H2N2|NGT_ACTP7 UDP-glucose:protein N-beta-glucosyltransferase OS=Actinobacillus pleuropneumoniae serotype 7 (strain AP76) GN=APP7_1697 PE=1 SV=1
MENENKPNVANFEAAVAVKDYEKACSELLLILSQLDSNFGGIQEIEFEYPVQLQDLEQEK
IVYFCTRMATAITTLFSDPVLEISDLGVQRFLVYQRWLALIFASSPFVNADHILQTYNRE
PNRKNSLEIHLDSSKSSLIKFCILYLPESNVNLNLDVMWNISPELCASLCFALQSPRFIG
TSTAFNKRATILQWFPRHLDQLKNLNNIPSAISHDVYMHCSYDTSVNKHDVKRALNHVIR
RHIESEYGWKDRYVAHIGYRNNKPVMVVLLEHFHSAHSIYRTHSTSMIAAREHFYLIGLG
SPSVDQAGQEVFDEFHLVAGDNMKQKLEFIRSVCESNGAAIFYMPSIGMDMTTIFASNTR
LAPIQAIALGHPATTHSDFIEYVIVEDDYVGSEACFSETLLRLPKDALPYVPSALAPEKV
DYLLRENPEVVNIGIASTTMKLNPYFLEALKAIRDRAKVKVHFHFALGQSNGITHPYVER
FIKSYLGDSATAHPHSPYHQYLRILHNCDMMVNPFPFGNTNGIIDMVTLGLVGVCKTGAE
VHEHIDEGLFKRLGLPEWLIANTVDEYVERAVRLAENHQERLELRRYIIENNGLNTLFTG
DPRPMGQVFLEKLNAFLKEN |
| EMBL CDS | ACE62349.1 |
| Sequence length | 620 AA |
| Subcellular Location | Cytoplasm |
| Potential Application | 1) ApNGT novel abilities could use a promising tool for the production of glycoconjugates and for site-specific modification of proteins. |
| Additional Information | 1) ApNGT is metal independent glycosyl transfer as it glycosylated the peptide in the presence of EDTA.
2) The N-linked glucose can be elongated by a polymerizing alpha6GlcT, which is sensitive to the UDP-glucose concentration.
3) ApNGT shows a relaxed peptide substrate specificity in vivo. In addition to the canonical NX(S/T) glycosylation consensus sites, it also glycosylates asparagine residues in NXA, NXG, NXD, and NXV sequons. |
| Glycosyltransferase Information |
| Glycosylation Type | N- (Asn) linked |
| CAZY Family | GT41 |
| EC Number (BRENDA) | 2.4.1.- |
| Mechanism of Glycan Transfer | Sequential |
| Acceptor specificity Sequon_1 | NX(S/T) |
| Donor Type | Nucleotide activated sugars |
| Donor Specificity | UDP-Glc and UDP-Gal ( UDP-Glc is preferred over UDP-Gal) |
| Accessory GT ID | ProGT42.1 |
| Glycan Information |
| Glycan transferred | Monosaccharides (Glc and Gal)Â |
| Method of Glycan Indentification | NanoLC-ESI-MS/MS (HCD and ETD), MS and MS/MS, NMR and MALDI-TOF MS |
| Experimental_strategies | In vitro and In vivo  |
| Acceptor Subtrate Information |
| Acceptor Substrate name | Hexapeptide DANYTK (labeled with TAMRA at the N terminus) |
| Acceptor Substrate name | Synthetic Acceptor Peptide of Yeast glycoprotein lysophospholipase 2 (Plb2): SIVNPGGSNLTYIER |
| Acceptor Substrate name | AcrA |
| ProGPdb ID | ProGP217 |
| Acceptor Substrate name | Autotransporter adhesin |
| ProGPdb ID | ProGP450 |
| Acceptor Substrate name | AtaC1866–2428 |
| ProGPdb ID | ProGP450 |
| Acceptor Substrate name | COK_1394-62–640 |
| ProGPdb ID | ProGP451 |
| Acceptor Substrate name | HMW1ct |
| ProGPdb ID | ProGP227 |
| Acceptor Substrate name | human erythropoietin (hEPO) |
| Acceptor Substrate name | Cholera toxin subunit B (CtxB) |
| Acceptor Substrate name | GAPDH |
| Acceptor Substrate name | PCKA |
| Acceptor Substrate name | PurH |
| Acceptor Substrate name | ArgG |
| Acceptor Substrate name | DnaK (Chaperone protein) |
| Acceptor Substrate name | YuaO (Uncharacterized protein) |
| Acceptor Substrate name | MdtE( Multidrug resistance protein) |
| Acceptor Substrate name | ArgG(Argininosuccinate synthase) |
| Litrature |
| Year Of Validation | 2011Â |
| Reference | Schwarz, F., Fan, Y. Y., Schubert, M., & Aebi, M. (2011). Cytoplasmic N-glycosyltransferase of Actinobacillus pleuropneumoniae is an inverting enzyme and recognizes the NXS/T consensus sequence. Journal of Biological Chemistry, jbc-M111.
|
| Authors | Schwarz, F., Fan, Y. Y., Schubert, M., & Aebi, M.
|
| Research groups | Molecular Biology and Biophysics, Department of Biology, ETH Zürich, 8093 Zürich, Switzerland
|
| Corresponding Author | Aebi, M.
|
| Contacts | Molecular Biology and Biophysics, Department of Biology, ETH Zürich, 8093 Zürich, Switzerland
Inst. of Microbiology, Dept. of Biology, ETH Zürich, Wolfgang-Pauli-Str. 10, 8093 Zürich, Switzerland
|
| Reference | Naegeli, A., Michaud, G., Schubert, M., Lin, C.W., Lizak, C., Darbre, T., Reymond, J.L. & Aebi, M. (2014). Substrate specificity of cytoplasmic N-glycosyltransferase. Journal of biological chemistry, jbc-M114.
|
| Authors | Naegeli, A., Michaud, G., Schubert, M., Lin, C.W., Lizak, C., Darbre, T., Reymond, J.L. & Aebi, M.
|
| Research groups | 1 Department of Biology, Institute of Microbiology, ETH Zurich, CH-8093 Zurich.
2 Department of Chemistry and Biochemistry, University of Berne, 3012 Berne, and.
3 Department of Biology, Institute of Molecular Biology and Biophysics, ETH Zurich, CH-8093 Zurich, Switzerland.
4 Department of Biology, Institute of Microbiology, ETH Zurich, CH-8093 Zurich,
|
| Corresponding Author | Aebi, M.
|
| Contacts | 1 Department of Biology, Institute of Microbiology, ETH Zurich, CH-8093 Zurich.
|
| Reference | Naegeli A, Neupert C, Fan YY, Lin CW, Poljak K, Papini AM, Schwarz F, Aebi M. Molecular analysis of an alternative N-glycosylation machinery by functional transfer from Actinobacillus pleuropneumoniae to Escherichia coli. Journal of Biological Chemistry. 2013 Nov 25:jbc-M113.
|
| Authors | Naegeli A, Neupert C, Fan YY, Lin CW, Poljak K, Papini AM, Schwarz F, Aebi M.
|
| Research groups |
1 Institute of Microbiology, Swiss Federal Institute of Technology, ETH Zurich, CH-8093 Zurich, Switzerland
|
| Corresponding Author | Aebi M.
|
| Contacts |
1 Institute of Microbiology, Swiss Federal Institute of Technology, ETH Zurich, CH-8093 Zurich, Switzerland
|
