| ProGlyProt ID | BC140 |
| Organism Information |
| Organism Name | Escherichia coli K12 |
| 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 | flu |
| NCBI Gene ID | 946540 |
| GenBank Gene Sequence | 946540 |
| Protein Information |
| Protein Name | Ag43α (Antigen 43 passenger domain) |
| UniProtKB/SwissProt ID | P39180 |
| NCBI RefSeq | YP_026164.1 |
| EMBL-CDS | AAT48141.1 |
| UniProtKB Sequence | >sp|P39180|AG43_ECOLI Antigen 43 OS=Escherichia coli (strain K12) GN=flu PE=1 SV=3
MKRHLNTCYRLVWNHMTGAFVVASELARARGKRGGVAVALSLAAVTSLPVLAADIVVHPG
ETVNGGTLANHDNQIVFGTTNGMTISTGLEYGPDNEANTGGQWVQDGGTANKTTVTSGGL
QRVNPGGSVSDTVISAGGGQSLQGRAVNTTLNGGEQWMHEGAIATGTVINDKGWQVVKPG
TVATDTVVNTGAEGGPDAENGDTGQFVRGDAVRTTINKNGRQIVRAEGTANTTVVYAGGD
QTVHGHALDTTLNGGYQYVHNGGTASDTVVNSDGWQIVKNGGVAGNTTVNQKGRLQVDAG
GTATNVTLKQGGALVTSTAATVTGINRLGAFSVVEGKADNVVLENGGRLDVLTGHTATNT
RVDDGGTLDVRNGGTATTVSMGNGGVLLADSGAAVSGTRSDGKAFSIGGGQADALMLEKG
SSFTLNAGDTATDTTVNGGLFTARGGTLAGTTTLNNGAILTLSGKTVNNDTLTIREGDAL
LQGGSLTGNGSVEKSGSGTLTVSNTTLTQKAVNLNEGTLTLNDSTVTTDVIAQRGTALKL
TGSTVLNGAIDPTNVTLASGATWNIPDNATVQSVVDDLSHAGQIHFTSTRTGKFVPATLK
VKNLNGQNGTISLRVRPDMAQNNADRLVIDGGRATGKTILNLVNAGNSASGLATSGKGIQ
VVEAINGATTEEGAFVQGNRLQAGAFNYSLNRDSDESWYLRSENAYRAEVPLYASMLTQA
MDYDRIVAGSRSHQTGVNGENNSVRLSIQGGHLGHDNNGGIARGATPESSGSYGFVRLEG
DLMRTEVAGMSVTAGVYGAAGHSSVDVKDDDGSRAGTVRDDAGSLGGYLNLVHTSSGLWA
DIVAQGTRHSMKASSDNNDFRARGWGWLGSLETGLPFSITDNLMLEPQLQYTWQGLSLDD
GKDNAGYVKFGHGSAQHVRAGFRLGSHNDMTFGEGTSSRAPLRDSAKHSVSELPVNWWVQ
PSVIRTFSSRGDMRVGTSTAGSGMTFSPSQNGTSLDLQAGLEARVRENITLGVQAGYAHS
VSGSSAEGYNGQATLNVTF |
| Sequence length | 1039 AA |
| Subcellular Location | Secreted |
| Function | Self-associating autotransporters (SAAT). It is the extracellular passenger domain of the E. coli autotransporter which represents a branch of the type V secretion pathway. Mediates bacterial autoaggregation (Ag43 interacts with Ag43 intercellularly) and biofilm formation as well as adhesion to and invasion of mammalian cells. It causes frizzy colony morphology and cell settling. |
| Protein Structure |
| PDB ID | |
| Glycosylation Status |
| Glycosylation Type | O (Ser) linked |
| Experimentally Validated Glycosite(s) in Full Length Protein | (Signal pepide: 1-52; sequence beyond 551 is cleaved during secretion) T87, T113, S117, S135, T149, T150, T229, T232, T287/T288, S317, S332, S396, S406, S422, T487, S491, S503 |
| Experimentally Validated Glycosite(s ) in Mature Protein | T35, T61, S65, S83, T97, T98, T177, T180, T235/T236, S265, S280, S344, S354, S370, T435, S439, S451 |
| Glycosite(s) Annotated Protein Sequence | >sp|P39180|AG43_ECOLI Antigen 43 OS=Escherichia coli (strain K12) GN=flu PE=1 SV=3
MKRHLNTCYRLVWNHMTGAFVVASELARARGKRGGVAVALSLAAVTSLPVLAADIVVHPG
ETVNGGTLANHDNQIVFGTTNGMTIST*(87)GLEYGPDNEANTGGQWVQDGGTANKT*(113)TVTS*(117)GGL
QRVNPGGSVSDTVIS*(135)AGGGQSLQGRAVNT*(149)T(150)LNGGEQWMHEGAIATGTVINDKGWQVVKPG
TVATDTVVNTGAEGGPDAENGDTGQFVRGDAVRTTINKNGRQIVRAEGT*(229)ANT*(232)TVVYAGGD
QTVHGHALDTTLNGGYQYVHNGGTASDTVVNSDGWQIVKNGGVAGNT*(287)T*(288)VNQKGRLQVDAG
GTATNVTLKQGGALVTS*(317)TAATVTGINRLGAFS*(332)VVEGKADNVVLENGGRLDVLTGHTATNT
RVDDGGTLDVRNGGTATTVSMGNGGVLLADSGAAVS*(396)GTRSDGKAFS*(406)IGGGQADALMLEKG
SS*(422)FTLNAGDTATDTTVNGGLFTARGGTLAGTTTLNNGAILTLSGKTVNNDTLTIREGDAL
LQGGSLT*(487)GNGS*(491)VEKSGSGTLTVS*(503)NTTLTQKAVNLNEGTLTLNDSTVTTDVIAQRGTALKL
TGSTVLNGAIDPTNVTLASGATWNIPDNATVQSVVDDLSHAGQIHFTSTRTGKFVPATLK
VKNLNGQNGTISLRVRPDMAQNNADRLVIDGGRATGKTILNLVNAGNSASGLATSGKGIQ
VVEAINGATTEEGAFVQGNRLQAGAFNYSLNRDSDESWYLRSENAYRAEVPLYASMLTQA
MDYDRIVAGSRSHQTGVNGENNSVRLSIQGGHLGHDNNGGIARGATPESSGSYGFVRLEG
DLMRTEVAGMSVTAGVYGAAGHSSVDVKDDDGSRAGTVRDDAGSLGGYLNLVHTSSGLWA
DIVAQGTRHSMKASSDNNDFRARGWGWLGSLETGLPFSITDNLMLEPQLQYTWQGLSLDD
GKDNAGYVKFGHGSAQHVRAGFRLGSHNDMTFGEGTSSRAPLRDSAKHSVSELPVNWWVQ
PSVIRTFSSRGDMRVGTSTAGSGMTFSPSQNGTSLDLQAGLEARVRENITLGVQAGYAHS
VSGSSAEGYNGQATLNVTF
|
| Sequence Around Glycosites (21 AA) | FGTTNGMTISTGLEYGPDNEA
WVQDGGTANKTTVTSGGLQRV
GGTANKTTVTSGGLQRVNPGG
PGGSVSDTVISAGGGQSLQGR
GQSLQGRAVNTTLNGGEQWMH
QSLQGRAVNTTLNGGEQWMHE
NGRQIVRAEGTANTTVVYAGG
QIVRAEGTANTTVVYAGGDQT
IVKNGGVAGNTTVNQKGRLQV
VKNGGVAGNTTVNQKGRLQVD
TLKQGGALVTSTAATVTGINR
VTGINRLGAFSVVEGKADNVV
VLLADSGAAVSGTRSDGKAFS
SGTRSDGKAFSIGGGQADALM
ADALMLEKGSSFTLNAGDTAT
GDALLQGGSLTGNGSVEKSGS
LQGGSLTGNGSVEKSGSGTLT
EKSGSGTLTVSNTTLTQKAVN |
| Glycosite Sequence Logo | seqlogo |
| Glycosite Sequence Logo |  |
| Technique(s) used for Glycosylation Detection | Labelling with digoxigenin (DIG)-conjugated hydrazide. Subsequently, the DIG-labelled structures were detected with peroxidase-conjugated anti-DIG antibodies.. |
| Technique(s) used for Glycosylated Residue(s) Detection | Automated LC-ESI–MS/MS (liquid chromatography electrospray ionization tandem mass spectrometry) with CID (collision induced dissociation) and ETD (electron transfer dissociation) after β-elimination. |
| Protein Glycosylation- Implication | Glycosylation protects Ag43α against native-state proteolysis and stabilizes it against thermal and chemical denaturation, and also increases its refolding rate. It reduces the stabilizing effect of Ca2+ ions, prevents Ca2+ to promote cell adhesion and inhibits the bacterial amyloid-forming ability of Ag43α. |
| Glycan Information |
| Glycan Annotation | Multiple heptose residues. |
| Technique(s) used for Glycan Identification | |
| Protein Glycosylation linked (PGL) gene(s) |
| OST Gene Name | aah (autotransporter-adhesin-heptosyltransferase) charcaterized based on gene deletion studies |
| OST NCBI Gene ID | |
| OST GenBank Gene Sequence | |
| OST Protein Name | Aah is the heptosyltransferase. Tib C (E. coli (ET |
| 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) Knudsen, S.K., Stensballe, A., Franzmann, M., Westergaard, U.B. and Otzen, D.E. (2008) Effect of glycosylation on the extracellular domain of the Ag43 bacterial autotransporter: enhanced stability and reduced cellular aggregation. Biochem J, 412, 563-577. [PubMed: 18341480]
2) Sherlock, O., Dobrindt, U., Jensen, J.B., Munk Vejborg, R. and Klemm, P. (2006) Glycosylation of the self-recognizing Escherichia coli Ag43 autotransporter protein. J Bacteriol, 188, 1798-1807. [PubMed: 16484190] |
| Additional Comments | Ag43 protein is involved in phase variable biofilm formation.
The glycosylated peptides were identified in a region of the protein composed of imperfect 19-amino-acid repeats.
Glycosylation by heptoses represents a novel protein modification in eubacteria. |
| Year of Identification | 2006 |
| Year of Validation | 2008 |
