| 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 |
