ProGP249 (Ag43 α (Antigen 43 passenger domain))

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ProGP ID ProGP249 (Ag43 α (Antigen 43 passenger domain))
Validation Status Characterized
Organism Information
Organism NameEscherichia coli K12
Domain Bacteria
Taxonomic ID (NCBI) 83333
Genome Information
GenBank U00096.2
EMBL U00096
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. There are a myriad of serotypes of pathogenic E. coli. Adhesion to the host cells is an important step in its pathogenesis. However, most strains are harmless and normal flora residing in the gut.
Gene Information
Gene Nameflu
NCBI Gene ID 946540
GenBank Gene Sequence NC_000913.3
Protein Information
Protein NameAg43α (Antigen 43 passenger domain)
UniProtKB/SwissProt ID P39180
NCBI RefSeq YP_026164.1
EMBL-CDSAAT48141.1
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 LocationSecreted
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.
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 ProteinT35, 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
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Technique(s) used for Glycosylation DetectionLabelling 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 Ag43 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 Ag43.
Glycan Information
Glycan Annotation Multiple heptose residues.
Protein Glycosylation linked (PGL) gene(s)
OST Gene NameAah is the heptosyltransferase. Tib C (E. coli (ETEC) strain H10407) has sequence similarity with Aah gene and it can substitute function of Aah.
OST ProGT IDProGT8
Additional CommentAg43 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.
Literature
Year of Identification2006
Year of Identification Month Wise2006.3.1
Year of Validation 2008
ReferenceKnudsen, 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. Biochemical Journal, 412(3), pp.563-577.
Corresponding Author Daniel E. Otzen
ContactDepartment of Life Sciences, Aalborg University, Sohngaardsholmsvej 49, DK-9000 Aalborg, Denmark.
ReferenceSherlock, O., Dobrindt, U., Jensen, J.B., Munk Vejborg, R. and Klemm, P., 2006. Glycosylation of the self-recognizing Escherichia coli Ag43 autotransporter protein. Journal of bacteriology, 188(5), pp.1798-1807.
Corresponding Author Per Klemm
ContactCentre for Biomedical Microbiology, BioCentrum-DTU, Bldg. 301, Technical University of Denmark, DK-2800 Lyngby, Denmark.