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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)
 
 
Genome Sequence(s)
GenBank
EMBL
Organism Additional Information
Escherichia coli (Gram-negative) is the predominant facultative organism in the human intestine. It is responsible for a number of diseases like urinary tract infections, gastroenteritis (diarrhoea), meningitis, traveler's diarrhea and hemorrhagic colitis.
 
 
Gene Information
Gene Name
potD (b1123)
NCBI Gene ID
GenBank Gene Sequence
 
 
Protein Information
Protein Name
Spermidine/putrescine-binding protein (PotD)
UniProtKB/SwissProt ID
NCBI RefSeq
EMBL-CDS
UniProtKB Sequence
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
 
 
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=Esche
richia coli (strain K12) GN=potD PE=1 SV=1 MKKWSRHLLAAGALALGMSAAHADDN*(26)
NTLYFYNWTEYVPPGLLEQFTKETGIKVIYSTYE SN*(62)ETMYAKLKTYKDGAYDLVVPSTYYVDKMRK
EGMIQKIDKSKLTNFSNLDPDMLNKPFD PNNDYSIPYIWGATAIGVNGDAVDPKSVTSWADLWKPEYKGSLLLT
DDAREVFQMALRKL GYSGNTTDPKEIEAAYNELKKLMPNVAAFNSDNPANPYMEGEVNLGMIWNGSAFVARQAG
TPIDVVWPKEGGIFWMDSLAIPANAKNKEGALKLINFLLRPDVAKQVAETIGYPTPNLAA RKLLSPEVANDK
TLYPDAETIKNGEWQNDVGAASSIYEEYYQKLKAGR
Sequence Around Glycosites (21 AA)
LGMSAAHADDNNTLYFYNWTE
IKVIYSTYESNETMYAKLKTY
Glycosite Sequence Logo
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)
 
 
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).
 
 
Literature
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. [PubMed: 8897598]
2) Sugiyama, S., Vassylyev, D.G., Matsushima, M., Kashiwagi, K., Igarashi, K. and Morikawa, K. (1996) Crystal structure of PotD, the primary receptor of the polyamine transport system in Escherichia coli. J Biol Chem, 271, 9519-9525. [PubMed: 8621624]
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 lacking D at -2 position but with low efficiency.
UniProtKB ID for ClPglB is B9KDD4.
Year of Identification
2011
Year of Validation
2011
 
 
Copyright @ 2011 IMTECH
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