ProGlyProt ID BC173 Organism Information Organism Name Escherichia coli K12/DH5alpha Domain Bacteria Classification Family: Enterobacteriaceae Order: "Enterobacteriales" Class: Gammaproteobacteria Division or phylum: "Proteobacteria" Taxonomic ID (NCBI) 83333 Genome Sequence (s) GeneBank U00096.2 EMBL U00096 Gene Information Gene Name potD (b1123) NCBI Gene ID 945682 GenBank Gene Sequence 945682 Protein Information Protein Name Spermidine/putrescine-binding protein (PotD) UniProtKB/SwissProt ID P0AFK9 NCBI RefSeq NP_415641.1 EMBL-CDS AAC74207.1 UniProtKB Sequence >sp|P0AFK9|POTD_ECOLI Spermidine/putrescine-binding periplasmic protein OS=Escherichia coli (strain K12) GN=potD PE=1 SV=1 MKKWSRHLLAAGALALGMSAAHADDNNTLYFYNWTEYVPPGLLEQFTKETGIKVIYSTYE SNETMYAKLKTYKDGAYDLVVPSTYYVDKMRKEGMIQKIDKSKLTNFSNLDPDMLNKPFD PNNDYSIPYIWGATAIGVNGDAVDPKSVTSWADLWKPEYKGSLLLTDDAREVFQMALRKL GYSGNTTDPKEIEAAYNELKKLMPNVAAFNSDNPANPYMEGEVNLGMIWNGSAFVARQAG TPIDVVWPKEGGIFWMDSLAIPANAKNKEGALKLINFLLRPDVAKQVAETIGYPTPNLAA RKLLSPEVANDKTLYPDAETIKNGEWQNDVGAASSIYEEYYQKLKAGR Sequence length 348 AA Subcellular Location Periplasm Function It is a component of the polyamine transport system specifically binding either spermidine or putrescine. Protein Structure PDB ID 1POT, 1POY Glycosylation Status Glycosylation Type N (Asn) linked Experimentally Validated Glycosite(s) in Full Length Protein N26, N62 Experimentally Validated Glycosite(s ) in Mature Protein N26, N62 Glycosite(s) Annotated Protein Sequence >sp|P0AFK9|POTD_ECOLI Spermidine/putrescine-binding periplasmic protein OS=Escherichia coli (strain K12) GN=potD PE=1 SV=1 MKKWSRHLLAAGALALGMSAAHADDN*(26)NTLYFYNWTEYVPPGLLEQFTKETGIKVIYSTYE SN*(62)ETMYAKLKTYKDGAYDLVVPSTYYVDKMRKEGMIQKIDKSKLTNFSNLDPDMLNKPFD PNNDYSIPYIWGATAIGVNGDAVDPKSVTSWADLWKPEYKGSLLLTDDAREVFQMALRKL GYSGNTTDPKEIEAAYNELKKLMPNVAAFNSDNPANPYMEGEVNLGMIWNGSAFVARQAG TPIDVVWPKEGGIFWMDSLAIPANAKNKEGALKLINFLLRPDVAKQVAETIGYPTPNLAA RKLLSPEVANDKTLYPDAETIKNGEWQNDVGAASSIYEEYYQKLKAGR Sequence Around Glycosites (21 AA) LGMSAAHADDNNTLYFYNWTE IKVIYSTYESNETMYAKLKTY Glycosite Sequence Logo seqlogo Glycosite Sequence Logo Technique(s) used for Glycosylation Detection SBA (soybean agglutinin) lectin-agarose affinity chromatography Technique(s) used for Glycosylated Residue(s) Detection Site-directed mutagenesis (N26Q, N62Q) Protein Glycosylation- Implication Glycan Information Glycan Annotation Linkage: Bac-Asn. GalNAc-α-GalNAc-α-GalNAc-α-GalNAc-α-GalNAc-1,3-Bac, where Bac is bacillosamine, 2,4-diacetamido-2,4,6-trideoxyglucopyranose. Technique(s) used for Glycan Identification MALDI-MS/MS (matrix-assisted laser desorption/ionization tandem mass spectrometry). Protein Glycosylation linked (PGL) gene(s) OST Gene Name OST NCBI Gene ID OST GenBank Gene Sequence OST Protein Name OST UniProtKB/ SwissProt ID OST NCBI RefSeq OST EMBL-CDS OST UniProtKB Sequence OST EC Number (BRENDA) OST Genome Context Characterized Accessory Gene(s) PGL Additional Links CAZy Literatures Reference(s) 1) Schwarz, F., Lizak, C., Fan, Y.Y., Fleurkens, S., Kowarik, M. and Aebi, M. (2011) Relaxed acceptor site specificity of bacterial oligosaccharyltransferase in vivo. Glycobiology, 21, 45-54. [PubMed: 20847188] 2) Sugiyama, S., Matsuo, Y., Maenaka, K., Vassylyev, D.G., Matsushima, M., Kashiwagi, K., Igarashi, K. and Morikawa, K. (1996) The 1.8-A X-ray structure of the Escherichia coli PotD protein complexed with spermidine and the mechanism of polyamine binding. Protein Sci, 5, 1984-1990. [ Additional Comments Engineered glycoprotein. Native PotD protein is unglycosylated. It is glycosylated using Campylobacter lari glycosylation machinery (encoded by its pgl gene cluster) reconstituted in E. coli. Sequon features: The enzyme ClPglB has a predominant specificity for the sequon D/E-X'-N-X"-S/T (where X' and X" stand for any amino acid except proline) which is efficiently glycosylated. It has also a relaxed specificity toward sequences like DANSG and NNNST. Hence ClPglB can glycosylate sequons lack Year of Identification 2011 Year of Validation 2011