ProGlyProt ID BC144 Organism Information Organism Name Flavobacterium meningosepticum (Elizabethkingia meningoseptica) Domain Bacteria Classification Family: Flavobacteriaceae Order: "Flavobacteriales" Class: Flavobacteria Division or phylum: "Bacteroidetes" Taxonomic ID (NCBI) 238 Genome Sequence (s) EMBL L06332 Gene Information Gene Name endOF3 NCBI Gene ID Protein Information Protein Name Endo-β-N-acetylglucosaminidase F3 UniProtKB/SwissProt ID P36913 NCBI RefSeq EMBL-CDS AAA24924.1 UniProtKB Sequence >sp|P36913|EBA3_FLAME Endo-beta-N-acetylglucosaminidase F3 OS=Flavobacterium meningosepticum GN=endOF3 PE=1 SV=1 MKKIFFAQCSILLLMLGSCSKMTEDMTPESVNKEASVKSATALAGSNGVCIAYYITDGRN PTFKLKDIPDKVDMVILFGLKYWSLQDTTKLPGGTGMMGSFKSYKDLDTQIRSLQSRGIK VLQNIDDDVSWQSSKPGGFASAAAYGDAIKSIVIDKWKLDGISLDIEHSGAKPNPIPTFP GYAATGYNGWYSGSMAATPAFLNVISELTKYFGTTAPNNKQLQIASGIDVYAWNKIMENF RNNFNYIQLQSYGANVSRTQLMMNYATGTNKIPASKMVFGAYAEGGTNQANDVEVAKWTP TQGAKGGMMIYTYNSNVSYANAVRDAVKN Sequence length 329 AA Subcellular Location Periplasm (secreted) Function Endohydrolysis of the di-N-acetylchitobiosyl unit in high-mannose glycopeptides and glycoproteins. Hydrolyzes bi- and triantennary glycans. The presence of a core-bound fucose greatly augments endo F3 activity on biantennary and, presumably, triantennary oligosaccharides. EC= 3.2.1.96. Protein Structure PDB ID 1EOK, 1EOM Glycosylation Status Glycosylation Type O (Thr) linked Experimentally Validated Glycosite(s) in Full Length Protein (Signal peptide: 1-39) T88 Experimentally Validated Glycosite(s ) in Mature Protein T49 Glycosite(s) Annotated Protein Sequence >sp|P36913|EBA3_FLAME Endo-beta-N-acetylglucosaminidase F3 OS=Flavobacterium meningosepticum GN=endOF3 PE=1 SV=1 MKKIFFAQCSILLLMLGSCSKMTEDMTPESVNKEASVKSATALAGSNGVCIAYYITDGRN PTFKLKDIPDKVDMVILFGLKYWSLQDT*(88)TKLPGGTGMMGSFKSYKDLDTQIRSLQSRGIK VLQNIDDDVSWQSSKPGGFASAAAYGDAIKSIVIDKWKLDGISLDIEHSGAKPNPIPTFP GYAATGYNGWYSGSMAATPAFLNVISELTKYFGTTAPNNKQLQIASGIDVYAWNKIMENF RNNFNYIQLQSYGANVSRTQLMMNYATGTNKIPASKMVFGAYAEGGTNQANDVEVAKWTP TQGAKGGMMIYTYNSNVSYANAVRDAVKN Sequence Around Glycosites (21 AA) FGLKYWSLQDTTKLPGGTGMM Glycosite Sequence Logo seqlogo Glycosite Sequence Logo Technique(s) used for Glycosylation Detection Mass shift detected on SDS-polyacrylamide gel and TFA (trifluoroacetic acid) hydrolysis of intact glycopeptide followed by HPAEC chromatography on a PA-1 column Technique(s) used for Glycosylated Residue(s) Detection Edman degradation and collision activated dissociation (CAD) mass spectrometry Protein Glycosylation- Implication Glycosylation may contribute towards protein stability Glycan Information Glycan Annotation Linkage: Man- Thr. Branched, acidic, heptasaccharide (1244 Da) containing 3 different uronyl analogs, a methylated Rham and Man, a Glc, and a reducing terminal Man. Only pyranose ring forms were detected [(2-OMe)Man1-4GlcNAcU1-4GlcU1-4Glc1-4(2-OMe)GlcU-4[(2-OMe)Rham1-2]Man]. Technique(s) used for Glycan Identification ESI-MS (electrospray ionization mass spectrometry), CID (collision-induced dissociation), and a combination of isotopic labeling, composition and methylation analysis. 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) Waddling, C.A., Plummer, T.H., Jr., Tarentino, A.L. and Van Roey, P. (2000) Structural basis for the substrate specificity of endo-beta-N-acetylglucosaminidase F(3). Biochemistry, 39, 7878-7885. [PubMed: 10891067] 2) Plummer, T.H., Jr., Tarentino, A.L. and Hauer, C.R. (1995) Novel, specific O-glycosylation of secreted Flavobacterium meningosepticum proteins. Asp-Ser and Asp-Thr-Thr consensus sites. J Biol Chem, 270, 13192-13196. [PubMed: 7768916] 3) Reinhold, B.B., Hauer, C.R., Plum Additional Comments Identified glycosylation Sequon features: Consensus sequon observed Asp-Ser (DS) or Asp-Thr-Thr (DTT), at turns. DT alone may not serve as a sequon. Interestingly, the experimentally examined other five nonglycosylated DT sequons were followed by N, D and Q. Post translational modification of 1.2-kDa was detected by MS in 1993 (Ref. no. 4). The protein migrated slowly on SDS-PAGE (compared to its theoretical weight) and was found to be heterogeneous by MS. Year of Identification 1995 Year of Validation 1995