ProGlyProt ID AC111 Organism Information Organism Name Methanococcus maripaludis S2 Domain Archaea Classification Family: Methanococcaceae Order: Methanococcales Class: Methanococci or Methanothermea Division or phylum: "Euryarchaeota" Taxonomic ID (NCBI) 39152 Genome Sequence (s) GeneBank BX950229.1 EMBL BX950229 Gene Information Gene Name flaB3 (MMP1668) NCBI Gene ID 2761414 GenBank Gene Sequence 2761414 Protein Information Protein Name FlaB3 UniProtKB/SwissProt ID Q6LWP1 NCBI RefSeq NP_988788.1 EMBL-CDS CAF31224.1 UniProtKB Sequence >tr|Q6LWP1|Q6LWP1_METMP Flagellin B3 OS=Methanococcus maripaludis GN=flaB3 PE=3 SV=1 MVKKFMKSKKGAVGIGTLIIFIAMVLVAAIAASVIINTAGKLQHKASTVGEESTEQVASG IQVLKVIGHADTKTTIDKLAVMVAPNVGGEIDLSTTILTLGTGDAKYSLVYDSTQHNADV KDDGSDSIFDETEWGSGSKYGLIVLQDSDNSTAEATHPTINYGDKVYITVTMDVNSTSKV SGEVIPDYGASGIVEFRAPSVFTETVVTLQ Sequence length 210 AA Subcellular Location Surface associated Function Flagellin Protein Structure PDB ID Glycosylation Status Glycosylation Type N (Asn) linked Experimentally Validated Glycosite(s) in Full Length Protein (Signal peptide: 1-11) N150, N175 Experimentally Validated Glycosite(s ) in Mature Protein N139, N164 Glycosite(s) Annotated Protein Sequence >tr|Q6LWP1|Q6LWP1_METMP Flagellin B3 OS=Methanococcus maripaludis GN=flaB3 PE=3 SV=1 MVKKFMKSKKGAVGIGTLIIFIAMVLVAAIAASVIINTAGKLQHKASTVGEESTEQVASG IQVLKVIGHADTKTTIDKLAVMVAPNVGGEIDLSTTILTLGTGDAKYSLVYDSTQHNADV KDDGSDSIFDETEWGSGSKYGLIVLQDSDN*(150)STAEATHPTINYGDKVYITVTMDVN*(175)STSKV SGEVIPDYGASGIVEFRAPSVFTETVVTLQ Sequence Around Glycosites (21 AA) YGLIVLQDSDNSTAEATHPTI KVYITVTMDVNSTSKVSGEVI Glycosite Sequence Logo seqlogo Glycosite Sequence Logo Technique(s) used for Glycosylation Detection Aberrant SDS-PAGE migration and glycan detection by MS Technique(s) used for Glycosylated Residue(s) Detection NanoLC–MS/MS (nano-liquid chromatography-electrospray tandem mass spectrometry) Protein Glycosylation- Implication Glycan is necessary for proper flagellar filament assembly and function. A minimal portion of the glycan (at least two sugars) is required to facilitate flagellar assembly and the presence of each additional sugar results in increased motility, as measured on semi-swarm plates. Glycan Information Glycan Annotation Linkage: β-GalNAc-Asn. N-linked tetrasaccharide (1036.4 Da) composed of N-acetylgalactosamine, di-acetylated glucuronic acid, an acetylated and acetamidino-modified mannuronic acid linked to threonine, and a novel terminal sugar: Sug-4-β-ManNAc3NAmA6Thr-4-β-GlcNAc3NAcA-3-β-GalNAc-Asn where Sug is a novel monosaccharide unit, (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-α-L-erythro-hexos-5-ulo-1,5-pyranose. Technique(s) used for Glycan Identification NanoLC–MS/MS analysis and 1H, 13C NMR spectroscopy Protein Glycosylation linked (PGL) gene(s) OST Gene Name aglB OST NCBI Gene ID 2761329 OST GenBank Gene Sequence 2761329 OST Protein Name AglB/STT3 subunit OST UniProtKB/ SwissProt ID Q6LXC8 OST NCBI RefSeq NP_988544.1 OST EMBL-CDS CAF30980.1 OST UniProtKB Sequence >tr|Q6LXC8|Q6LXC8_METMP Oligosaccharyl transferase, STT3 subunit OS=Methanococcus maripaludis GN=MMP1424 PE=4 SV=1 MGEFLNKVSDFFKKNEKIKIILILLFIGMMSFQIRAQTADMAFTDNSYLQDMFSDDNGRM YLTALDPYYYLRMTENYVNNDYSNVGETTVGIDGENIPYDTIQYAPPGREAGLVSALSIA TVLVYSVWNSIDSTVTIMNAAFWVPAIMSIFLGIPVFFIVRRNTASNIGGLVGALLLISS PSLLYKTSAGFSDTPIFEILPLLFIVWMIMEAIHEQENSKKSGIFGGIAAILIGLYPMMW SGWWYAFDITAGFLVLYTAYEYLTKSKNLKNVITTSLITLVGGAILVSLSTGLSGFINWI LSPIGFTVINEATKITGWPNVYMTVSELAIPTVTDIIENSVGNIWLLIAGISGILLSFVS FKHDKQKIDIKYALYLTLWLIATVYAATKGIRFVALMTPALAIGIGIFAGQIENIIKRYE KKVEYILYPVIGILSVITLIKYGGELFNILVPTTYVPIAVYLSIIALLVLAVYKIIDIIS EKEQAVKKVFGILLAFMLVFPSMAAAVPFYTAPTMNNGWMDSLSWIKSETPENSVVTCWW DNGHIYTWATRKMVTFDGGSQNTPRAYWVGHAFSTSDENLSVGILRMLATSGDSAYDDDS ILIKKTGSIKDTVDILNKILPLTRTEAKASLVNNYDLTDAEAEEVLDLTHPKVTNPDYLI TYNRMTSIASVWSMFGNWNFSLPASTENSDREMGYYQQLGGSAQDINGTTVVYIPLQETD SYRVINILEITDSEIKSANAVIDSNNQTSMQSPNFHKLILKVNGNVYEQETNENGDYSEI VRLEKLSDGTYQVYAWVSSKNLEDSIYTKLHFLDGYGLEKISLEKESVDPTSYGIQPGFK VYSVDYGTDYLN OST EC Number (BRENDA) 2.4.1.119 OST Genome Context Q6LXC8 Characterized Accessory Gene(s) AglO, AglA, AglL are glycosyltransferases responsible for tetrasaccharide assembly. AglO adds the second sugar, AglA adds the third sugar, and AglL transfers the last sugar. MMP0350 (acetyltransferase-second sugar ), MMP1081 (Acetamidino transfer-third sugar), MMP1085 (Methyltransferase-fourth sugar). MMP1081 and MMP1085 are unpublished results as referred in ref. 4. Characterized by gene deletion studies. PGL Additional Links CAZy Literatures Reference(s) 1) Jarrell, K.F., Jones, G.M. and Nair, D.B. Biosynthesis and role of N-linked glycosylation in cell surface structures of archaea with a focus on flagella and s layers. Int J Microbiol, 2010, 470138. [PubMed: 20721273] 2) Kelly, J., Logan, S.M., Jarrell, K.F., VanDyke, D.J. and Vinogradov, E. (2009) A novel N-linked flagellar glycan from Methanococcus maripaludis. Carbohydr Res, 344, 648-653. [PubMed: 19203750] 3) VanDyke, D.J., Wu, J., Logan, S.M., Kelly, J.F., Mizuno, S., Aizawa, S Additional Comments Sug, a novel monosaccharide unit, (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-α-L-erythro-hexos-5-ulo-1,5-pyranose is the first example of a naturally occuring diglycoside of an aldulose. Year of Identification 2008 Year of Validation 2009