ProGlyProt ID BC139 Organism Information Organism Name Escherichia coli 278 Domain Bacteria Classification Family: Enterobacteriaceae Order: "Enterobacteriales" Class: Gammaproteobacteria Division or phylum: "Proteobacteria" Taxonomic ID (NCBI) 562 Genome Sequence (s) EMBL GU810159 Gene Information Gene Name aidA (plasmid encoded) NCBI Gene ID Protein Information Protein Name Diffuse Adherence Adhesin (AIDA-I) UniProtKB/SwissProt ID D7PPP4 NCBI RefSeq EMBL-CDS ADH10230.1 UniProtKB Sequence >tr|D7PPP4|D7PPP4_ECOLX Diffuse adherence adhesin OS=Escherichia coli GN=aidA PE=4 SV=1 MNKAYSIIWSHSRQAWIVASELARGHGFVLAKNTLLVLAVVSTIGNAFAVNISGTVSSGG TVSSGETQIVYSGGETSNATVNSGGTQIVNNGGKTTATTVNSSGSQNVGTSGATISTIVN SGGIQRVSSGGVASATNLSGGAQNIYNLGHASNTVIFSGGNQTIFSGGITDSTNISSGGQ QRVSSGGVASNTTINSSGAQNILSGGSAISTHISSGGNQYISAGANATETIVNSGGFQRV NSGAVATGTVLSGGTQNVSSGGSAISTSVYNSGVQTVFAGATVTDTTVNSGGNQNISSGG IVSETTVNVSGTQNIYSGGSALSANIKGSQIVNSEGTAINTLVSDGGYQHIRNGGIASGT IVNQSGYVNISSGGYAESTIINSGGTLRVLSDGYARGTILNNSGRENVSNGGVSYNAMIN TGGNQYIYSDGEATAAIVNTSGFQRINSGGTASGTKLSGGNQNVSSGGKAIDAEVYSGGK QTVYSGGEVSGTQIFNGGMVNVSGGTASGANVNLSGRLNAFAGNVVGTILNQEGRQYVYS GATATSTVGNNEGREYVLSGGITDGTVLNSGGLQAVSSGGKASATVINEGGAQFVYDGGQ VTGTNIKNGGTIRVDSGASALNIALSSGGNLFTSTGATVTGTNHYGSFSVSQNHASNIVL ENGGLLAVTSGGTATDTTVNSAGRLRIDDGGTINGTTTINADGIVAGTNIQNDGNFILNL AENYDFETELSGSGVLVKDNTGIMTYAGTLTQAQGVNVKNGGIIFDSAVVNADMAVNQNA YINISDQATINGSVNNNGSIVINNSIINGNITNDADLSFGTAKLLSATVNGSLVNNKNII LNPTKESAGNTLTVSNYTGTPGSVISLGGVLEGDNSLTDRLVVKGNTSGQSDIVYVNEDG SGGQTRDGINIISVEGNSDAEFSLKNRVVAGAYDYTLQKGNESGTDNKGWYLTSHLPTSD TRQYRPENGSYATNMALANSLFLMDLNERKQFRAMSDNTQPESASVWMKITGGRSSGKLN DGQNKTTTNQFINQLGGDIYKFHAEQLGDFTLGIMGGYANAKGKTINYTSNKAARNTLDG YSVGVYGTWYQNGENATGLFAETWMQYNWFNASVKGDGLEEEKYNLNGLTASAGGGYNLN VHTWTSPEGITGEFWLQPHLQAVWMGVTPDTHQEDNGTVVQGAGKNNIQTKAGIRASWKV KSTLDKDTGREFRPYIEANWIHNTHEFGVKMSDDSQLLSGSRNQGEIKTGIEGVITQNLS VNGGVAYQAGGHGSNAISGALGIKYSF Sequence length 1287 AA Subcellular Location Periplasm (Extracellular membrane associated) Function Self-associating autotransporters (SAAT). Adhesin involved in diffuse adherence. Able to confer a pattern of diffuse adherence on the surface of cultured epithelial cells. Protein Structure PDB ID Glycosylation Status Glycosylation Type O (Ser) linked Experimentally Validated Glycosite(s) in Full Length Protein S102, S111, S116, S242, S252, S334, S391, S409, S540, S546, S559, S570, S577, S578, S583, T154 Experimentally Validated Glycosite(s ) in Mature Protein S102, S111, S116, S242, S252, S334, S391, S409, S540, S546, S559, S570, S577, S578, S583, T154 Glycosite(s) Annotated Protein Sequence >tr|D7PPP4|D7PPP4_ECOLX Diffuse adherence adhesin OS=Escherichia coli GN=aidA PE=4 SV=1 MNKAYSIIWSHSRQAWIVASELARGHGFVLAKNTLLVLAVVSTIGNAFAVNISGTVSSGG TVSSGETQIVYSGGETSNATVNSGGTQIVNNGGKTTATTVNS*(102)SGSQNVGTS*(111)GATIS*(116)TIVN SGGIQRVSSGGVASATNLSGGAQNIYNLGHASNT*(154)VIFSGGNQTIFSGGITDSTNISSGGQ QRVSSGGVASNTTINSSGAQNILSGGSAISTHISSGGNQYISAGANATETIVNSGGFQRV NS*(242)GAVATGTVLS*(252)GGTQNVSSGGSAISTSVYNSGVQTVFAGATVTDTTVNSGGNQNISSGG IVSETTVNVSGTQNIYSGGSALSANIKGSQIVNS*(334)EGTAINTLVSDGGYQHIRNGGIASGT IVNQSGYVNISSGGYAESTIINSGGTLRVLS*(391)DGYARGTILNNSGRENVS*(409)NGGVSYNAMIN TGGNQYIYSDGEATAAIVNTSGFQRINSGGTASGTKLSGGNQNVSSGGKAIDAEVYSGGK QTVYSGGEVSGTQIFNGGMVNVSGGTASGANVNLSGRLNAFAGNVVGTILNQEGRQYVYS*(540)GATATS*(546)TVGNNEGREYVLS*(559)GGITDGTVLNS*(570) GGLQAVS*(577)S*(578) GGKAS*(583)ATVINEGGAQFVYDGGQ VTGTNIKNGGTIRVDSGASALNIALSSGGNLFTSTGATVTGTNHYGSFSVSQNHASNIVL ENGGLLAVTSGGTATDTTVNSAGRLRIDDGGTINGTTTINADGIVAGTNIQNDGNFILNL AENYDFETELSGSGVLVKDNTGIMTYAGTLTQAQGVNVKNGGIIFDSAVVNADMAVNQNA YINISDQATINGSVNNNGSIVINNSIINGNITNDADLSFGTAKLLSATVNGSLVNNKNII LNPTKESAGNTLTVSNYTGTPGSVISLGGVLEGDNSLTDRLVVKGNTSGQSDIVYVNEDG SGGQTRDGINIISVEGNSDAEFSLKNRVVAGAYDYTLQKGNESGTDNKGWYLTSHLPTSD TRQYRPENGSYATNMALANSLFLMDLNERKQFRAMSDNTQPESASVWMKITGGRSSGKLN DGQNKTTTNQFINQLGGDIYKFHAEQLGDFTLGIMGGYANAKGKTINYTSNKAARNTLDG YSVGVYGTWYQNGENATGLFAETWMQYNWFNASVKGDGLEEEKYNLNGLTASAGGGYNLN VHTWTSPEGITGEFWLQPHLQAVWMGVTPDTHQEDNGTVVQGAGKNNIQTKAGIRASWKV KSTLDKDTGREFRPYIEANWIHNTHEFGVKMSDDSQLLSGSRNQGEIKTGIEGVITQNLS VNGGVAYQAGGHGSNAISGALGIKYSF Sequence Around Glycosites (21 AA) GGKTTATTVNSSGSQNVGTSG NSSGSQNVGTSGATISTIVNS QNVGTSGATISTIVNSGGIQR VNSGGFQRVNSGAVATGTVLS SGAVATGTVLSGGTQNVSSGG ANIKGSQIVNSEGTAINTLVS INSGGTLRVLSDGYARGTILN ILNNSGRENVSNGGVSYNAMI LNQEGRQYVYSGATATSTVGN QYVYSGATATSTVGNNEGREY GNNEGREYVLSGGITDGTVLN GGITDGTVLNSGGLQAVSSGG VLNSGGLQAVSSGGKASATVI LNSGGLQAVSSGGKASATVIN LQAVSSGGKASATVINEGGAQ NIYNLGHASNTVIFSGGNQTI Glycosite Sequence Logo seqlogo Glycosite Sequence Logo Technique(s) used for Glycosylation Detection Aberrant mobility on SDS-polyacrylamide gel, labelling with digoxigenin (DIG) hydrazide after periodate oxidation. The glycan was not detectable by common lectins and could not be deglycosylated by a broad spectrum of glycosidases. Technique(s) used for Glycosylated Residue(s) Detection MS-MS (tandem mass spectrometry) and N-terminal sequencing of AIDA-I peptides Protein Glycosylation- Implication Glycosylation is required to ensure a normal conformation of AIDA-I. Glycans could be involved in receptor recognition making it essential for adhesion but not for autoaggregation or biofilm formation. It also provides increased resistance to degradation. Only the glycosylated form binds to cultured epithelial cells. Further unglycosylated AIDA-I is expressed in smaller amounts than its relatively thermostable glycosylated form. Unglycosylated AIDA-I is more sensitive to proteases and induces im Glycan Information Glycan Annotation Heterogenous multiple heptose residues (atleast 19 per protein molecule). Technique(s) used for Glycan Identification Gas chromatography and mass spectrometry after methanolysis and carbohydrate labelling. 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) Charbonneau, M.E., Girard, V., Nikolakakis, A., Campos, M., Berthiaume, F., Dumas, F., Lepine, F. and Mourez, M. (2007) O-linked glycosylation ensures the normal conformation of the autotransporter adhesin involved in diffuse adherence. J Bacteriol, 189, 8880-8889. [PubMed: 17951390] 2) Sherlock, O., Schembri, M.A., Reisner, A. and Klemm, P. (2004) Novel roles for the AIDA adhesin from diarrheagenic Escherichia coli: cell aggregation and biofilm formation. J Bacteriol, 186, 8058-8065. [P Additional Comments AIDA- I is primarily associated with pathogenic E. coli strains involved in neonatal and postweaning diarrhea in piglets. 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. Sequon features: The N-terminal repeats of AIDA-I that contain multiple copies of the consensus sequence VXNSGG, might serve as acceptor sites for the AAH heptosyltran Year of Identification 2001 Year of Validation 2007